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www.ZINEXTS.com Page 1 Vr.5.1
MagPurix Extraction Kit Handbook
For automated purification of nucleic acids using the MagPurix 12 System
1. ZP02001 MagPurix Blood DNA Extraction Kit 200
2. ZP02002 MagPurix Blood DNA Extraction Kit 1200
3. ZP02003 MagPurix Viral Nucleic Acid Extraction Kit
4. ZP02004 MagPurix Tissue DNA Extraction Kit
5. ZP02005 MagPurix Cultured Cell DNA Extraction Kit
6. ZP02006 MagPurix Bacterial DNA Extraction Kit
7. ZP02007 MagPurix HPV DNA Extraction Kit for Swab Samples
8. ZP02008 MagPurix TB DNA Extraction Kit
9. ZP02009 MagPurix FFPE DNA Extraction Kit
10. ZP02010 MagPurix Forensic DNA Extraction Kit
11. ZP02011 MagPurix Viral/Pathogen Nucleic Acids Extraction Kit A
12. ZP02012 MagPurix Viral/Pathogen Nucleic Acids Extraction Kit B
13. ZP02013 MagPurix Viral RNA Extraction kit
14. ZP02014 MagPurix Plant DNA Extraction Kit
15. ZP02015 MagPurix Total RNA Extraction Kit
16. ZP02016 MagPurix Viral Nucleic Acid Extraction Kit 800
For in vitro diagnostic use
Version: 5.1
Rev. Date: 01 JAN, 2014
Manual No. Z3001
Welkang Limited Suite B, 29 Harley Street
LONDON, W1G 9QR,
U.K.
ZINEXTS LIFE SCIENCE CORP.
2F.-2, NO.122, Qiaohe Rd., Zhonghe
Dist., New Taipei City 235, Taiwan
(R.O.C.)
TEL: +886 2 22463579
FAX: +886 2 22438570
www.ZINEXTS.com Page 2 Vr.5.1
TABLE OF CONTENTS
Page
REAGENT KIT SELECTION GUIDE 3
INTRODUCTION 4
The Zinexts Nucleic Acid Purification Technology 4
PRODUCT INFORMATION 5
Product Use Limitation, Warranty, Guarantee and Technical Support 5 Safety Info. , Manufacturer Info 6 ZP02001 MagPurix Blood DNA Extraction Kit 200 7 ZP02002 MagPurix Blood DNA Extraction Kit 1200 15 ZP02003 MagPurix Viral Nucleic Acid Extraction Kit 20 ZP02004 MagPurix Tissue DNA Extraction Kit 26 ZP02005 MagPurix Cultured Cell DNA Extraction Kit 35 ZP02006 MagPurix Bacterial DNA Extraction Kit 40 ZP02007 MagPurix HPV DNA Extraction Kit for Swab Samples 49 ZP02008 MagPurix TB DNA Extraction Kit 54 ZP02009 MagPurix FFPE DNA Extraction Kit 60 ZP02010 MagPurix Forensic DNA Extraction Kit 66 ZP02011 MagPurix Viral/Pathogen Nucleic Acids Extraction Kit A 73 ZP02012 MagPurix Viral/Pathogen Nucleic Acids Extraction Kit B 78 ZP02013 MagPurix Viral RNA Extraction kit
ZP02014 MagPurix Plant DNA Extraction Kit
ZP02015 MagPurix Total RNA Extraction Kit
ZP02016 MagPurix Viral Nucleic Acid Extraction Kit 800
83
88
93
101
PROTOCOL OF EXTRACTION 105
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Reagent Kit Selection Guide
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Introduction The Zinexts Nucleic Acid Purification Technology
Introduction
Zinexts Life Science is specialized in developing advanced,
efficient and reliable technologies in nucleic acid purification,
enabling successful delivery of extraction results from varied
sample types.
The MagPurix technology is a state of the art platform that
uses magnetic bead to extract nucleic acids from samples.
The platform commits a truly walk-away automation in
nucleic acid purification from samples to results. The
purification process contains steps of lysis, binding, washing
and elution as figure below.
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Product information
Intended use
MagPurix Kits is intended for the purification of nucleic acid
from biological specimen used with MagPurix 12s instrument
as IVD accessory.
The nucleic acids purified using the MagPurix 12s instrument
and reagent kits are suitable for a variety of polymerase chain
reaction (PCR) tests for human in vitro diagnostics purpose.
The MagPurix 12 instrument and reagent kits is intended for
professional use only.
Product Use
Limitations
The MagPurix instrument and MagPurix kit are not intended
for use as part of a specific in vitro diagnostic test. The user is
responsible for establishing performance characteristics
necessary for downstream diagnostic applications.
Appropriate controls must be included in any downstream
diagnostic applications using nucleic acid purified using the
MagPurix instrument and MagPurix reagent kits.
Warranty
Zinexts is committed to providing our customers with
high-quality products and services. Our goal is to ensure that
every customer is 100% satisfied with our products and our
service. If you have any questions or concerns about our
product or service, contact our Technical Support
Representatives.
Zinexts guarantees the performance of all products according
to specifications stated on our product literature. The
purchaser/user must determine the suitability of the product
for its particular use. We reserve the right to change, alter, or
modify any product to enhance its performance and design.
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This warranty limits Zinexts Life Science Corporation’s liability
only to the cost of the product. No warranty is granted for
products beyond their listed expiration date. No warranty is
applicable unless all product components are stored in
accordance with instructions.
Satisfaction
Guarantee
For any product that fails to perform satisfactorily due to any
reason other than misuse, Zinexts will replace it free of
charge. Simply call your distributor to get a replacement.
Technical
Support
For technical assistance and more information, please visit
our website at www.zinexts.com or call one of the Zinexts
Technical Service Departments or local distributors.
Safety
Information
When working with chemicals or samples, always wear a
suitable lab coat, disposable gloves, and protective goggles.
For more information, please consult the appropriate material
safety data sheets (MSDSs). You can find, download, view, and
print them from our website www.zinexts.com
Manufacturer
Information
Manufacturer:
Zinexts Life Science Corp.
Address:
2F.-2, No.122, Qiaohe Rd., Zhonghe Dist.,New Taipei City 235,
Taiwan
Tel: +886 2 2246 3579
Fax: +886 2 2243 8570
Mail: [email protected]
Product of Origin: TAIWAN
www.ZINEXTS.com Page 7 Vr.5.1
Kit Contents ZP02001-36 ZP02001-48
Reagent Cartridge 36 pcs (6x6) 48 pcs (4x6x2)
Reaction Chamber 36 pcs (6x6) 48 pcs (4x6x2)
Tip Holder 36 pcs 48 pcs(24x2)
Filtered Tip 38 pcs 50 pcs(25x2)
Piercing Pin 38 pcs 50 pcs(25x2)
Sample Tube (2 mL) 38 pcs 50 pcs(25x2)
Elute Tube (1.5 mL) 38 pcs 50 pcs(25x2)
Barcode Paper 1 pc 1 pc
MagPurix Blood DNA Extraction Kit 200
Cat. No. ZP02001 Process Time: 50-55 minutes
Intended Use
MagPurix Blood DNA Extraction Kit is used with the
MagPurix 12 instrument for extraction of DNA from
100-400μl mammalian whole blood, suspension of
mammalian blood cells
Application
Nucleic acids extracted from MagPurix Blood DNA Extraction
kit can be used in a number of downstream application
including: PCR, qPCR, Sequencing(NGS), Microarray, RFLP,
Southern Blot Analysis
Number Of Tests 36 or 48 extractions
Kit Components
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Reagent Cartridge Content
well-1 Proteinase K solution 40 μl
well-2 Lysis Buffer 2 1000 μl
well-3 Binding Buffer 1 600 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 Elution Buffer 1 1000 μl
well-9 Elution Buffer 2 1000 μl
well-10 Empty
Storage
MagPurix Blood DNA Extraction Kit should be
stored at room temperature (15-25°C). Do not
freeze the reagent cartridges. The Kits are stable
for 12 months under the condition
Store the purified total nucleic acid at 4 °C
(short-term) or aliquot and store at –70°C
(long-term) before perform the downstream
analysis
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Starting Material
Sample type Whole blood, Buffy coat, Leukocyte concentration*
Target nucleic acid Total DNA (Genomic DNA, mitochondrial and/or viral
DNA)**
Sample volume 100-400l whole blood (WBC count less than 2x 104 cells/l) ;
100-400l leukocyte concentration (contain no more than 5x 106 cells) ;
100-400l buffy coat**
NOTE:
* For those samples which have low leukocyte count(less than 1x103
cells/l), concentrate the blood cells by centrifuge at 3000 r.p.m. for
15min under 4°C and take the leukocyte concentration for DNA
extraction is recommended
** If the WBCs no. of blood sample were more than 2 x104
cells/l,
using MagPurix blood DNA extraction kit 1200 or dilute the blood
sample with PBS is recommended (e.g. the whole blood sample from
lymphoma/myeloma patient or other granulocyte-rich blood
sample/buffy coat)
Controls/Optional
internal control#
Add controls /internal control in the extraction procedure if the
downstream analysis needed (# see Controls/internal control on page
14 )
Elute volume 50-300l
If the sample volume is less, add the appropriate
volume of PBS
MagPurix Blood DNA Extraction Kit has proven to work
for fresh or frozen blood samples collected in tubes
containing common anti-coagulants like EDTA, heparin*
and citrate (*The EDTA is recommended to use as
anticoagulation agent, while heparin have inhibit effects
on nucleic acid amplification reaction)
Using fresh whole blood sample (within 1 week) for
extraction is recommended, the total nucleic acid yield
and quality would be decreased by time
Using the concentrated buffy coat (purified and free of
blood cells), the MagPurix Culture Cell DNA extraction
kit (ZP02005) is recommended
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If the whole blood sample is granulocyte-rich (white
blood cell no. more than 2 X 105 cells/l), dilute blood
sample or extract the DNA by the MagPurix blood DNA
extraction kit 1200(ZP02002) is recommended
For short-term storage (up to 10 days), store the tubes
at 2–8°C. However, for applications requiring maximum
fragment size, such as Southern blotting, storage
purified DNA at 2–8°C for up to 3 days , as low levels of
DNA degradation will occur after this time
For long-term storage, store the tubes at –70°C.
In fact, the extracted product contains total nucleic acid
(DNA and RNA), the RNA is not the major product in
this kit (about 10%) and would be degraded soon. If the
RNA-free product is needed, add some RNase to the
eluate
Result
(1) Expected Purity and Yield DNA was purified by using the MagPurix Blood DNA extraction Kit 200
and the MagPurix Purification Instrument using five different blood samples.
The DNA concentration was measured using a NanoDrop® 2000
spectrophotometer . The range of DNA yield is 2-18g ( from the sample of
WBCs counts : 2-20x103 cells/l)
Sample material Volume/amount DNA Yield Purity
Whole Blood
(WBC no. is 1.8 x 103 /ml)
100l
200l
300l
400l
1-1.2g
2-2.1g
2.8 -3.1g
4 -4.3g
OD260/OD280 >=1.7
OD260/OD230 >=1.5
Whole Blood
(WBC no. is 4 x 103 /l)
100l
200l
300l
400l
1.6 - 2.1g
3.8 - 3.9g
5.1 - 5.2g
8.5- 8.8g
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Whole Blood
(WBC no. is 6.9 x 103
/l)
100l
200l
300l
400l
2.9 - 3.1g
5.8- 6.2g
8.2 -8.8g
11.9 - 12.5g
Whole Blood
(WBC no. is 10.9 x 103 /l)
100l
200l
300l
400l
4.3- 4.6g
8.7-9.1g
11.5-12.2g
16.6- 17.5g
K562 cells 6 x 105
cells
2 x 106 cells
9-9.6g
22-25g
(2) Integrity
The DNA was isolated in replicates of 12 from 200l human whole blood
taken from two different donors. Elution volume was set to 100l. Integrity
of DNA was shown by subjecting each eluate to TAE agarose gel
electrophoresis together with a suitable molecular weight marker. All
samples shown single band with molecular weight at least 20kb with no
smear
(3) Scalability A. The DNA was isolated from different whole blood samples (WBCs
count range 1.8 – 22 X 103 cells/l). The DNA yield (measured by
Nanodrop 2000 UV-Vis spectrophotometer) shows excellent scalability
in different volume (100, 200, 300 and 400l) extraction and WBCs
count of the samples
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15
16
17
18
19
20
21
0 100 200 300 400 500
Sample volume (l)
Cro
ssin
g p
oin
t (C
t)
B. DNA isolated from different amounts of whole blood samples was
amplified by real-time qPCR by using theβ -globin gene specific
primers. The resulting crossing points confirm the scalability on
extraction
R² = 0.9959
R² = 0.9997
R² = 0.9981
R² = 0.9913
0
50
100
150
200
250
300
350
0 5 10 15 20 25
100ul
200ul
300ul
400ul
white blood no. ( E+3/l )
DN
A y
ield
(
l)
R² = 0.9935
0
20
40
60
80
100
120
140
0 100 200 300 400 500
Sample volume (l)
DN
A y
ield
(
g)
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(4) Reproducibility
The DNA were isolated by MagPurix blood DNA extraction kit from twenty
whole blood samples. The β-globin gene was detected by real-time qPCR. This data
shows ultra high stability and reproducibility of MagPurix 12s nucleic acid
purification system.
(5) Stability The DNA were extracted from whole blood by MagPurix Blood DNA Extraction
kit 200. Real-time qPCR detection of the β-globin (A) and spiked parvovirus B-19
DNA (A, B). No significant influences on samples with different anticoagulants and
storage conditions
(6) Sensitivity Using whole blood (in EDTA collection tube) spiked with serial-diluted human
Parvo B19 Virus (in range of 25-2500000 IU/ml). 200l sample were extracted and
eluted in 100l. 10l eluate was used for real-time PCR reaction by RealStar®
Parvovirus B19 PCR kit 1.0. As little of 5 IU spiked (about 1 IU in PCR reaction)
sample can be detected, proving the excellent sensitivity and linearity of isolation
procedure
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Controls/ internal control
Using appropriate controls for downstream analysis: :
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control (IC) Using a defined quantity control Place in sample tube or the round
well of the reaction chamber
Quality Control
In accordance with ZINEXTS’s ISO-certified Quality Management System, each lot of
MagPurix Blood DNA Extraction Kits is tested against predetermined specifications
to ensure consistent product quality
15
17
19
21
23
25
27
29
31
1 10 100 1000 10000 100000 1000000
Cro
ssin
g p
oin
t (C
t)
Human Parvo B19 virus DNA (IU/l)
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MagPurix Blood DNA Extraction Kit 1200
Cat. No. ZP02002 Process Time: 60-75 minutes
Intended Use
MagPurix Blood DNA Extraction Kit is used with the
MagPurix 12 instrument for extraction of gDNA from
400-1000μl mammalian blood, suspension of mammalian
blood cells.
Application
Nucleic acids extracted from MagPurix Blood DNA
Extraction kit can be used in a number of downstream
application including: PCR, qPCR, Sequencing(NGS),
Microarray, RFLP, Southern Blot Analysis
Number Of Tests
36 or 48 extractions
Kit Components
Kit Contents ZP02002-36 ZP02002-48
Reagent Cartridge 36 pcs (6x6) 48 pcs (4x6x2)
Reaction Chamber 36 pcs (6x6) 48 pcs (4x6x2)
Tip Holder 36 pcs 48 pcs(24x2)
Filtered Tip 38 pcs 50 pcs(25x2)
Piercing Pin 38 pcs 50 pcs(25x2)
Sample Tube (2 mL) 38 pcs 50 pcs(25x2)
Elute Tube (1.5 mL) 38 pcs 50 pcs(25x2)
Barcode Paper 1 pc 1 pc
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well-1 Proteinase K solution 50 μl
well-2 Lysis Buffer 2 1500 μl
well-3 Binding Buffer 1 1000 μl
well-4 Magnetic Bead Solution 1000 μl
well-5 Washing Buffer 1 1500 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 Elution Buffer 1 1000 μl
well-9 Elution Buffer 2 1000 μl
well-10 Empty
Reagent Cartridge Content
Storage
MagPurix Blood DNA Extraction Kit should be
stored at room temperature (15-25°C). Do not
freeze the reagent cartridges. The Kits are
stable for 12 months under the condition
Store the purified total nucleic acid at 4 °C
(short- term) or aliquot and store at -70°C
(long-term) before perform the downstream
analysis
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Starting Material Sample type Large volume whole blood sample, Buffy coat
Target nucleic acid Total DNA (Genomic DNA, mitochondrial and viral DNA)*
Sample volume 400-1000l whole blood (WBC count less than 6x 104cells/l) ;
400-1000l buffy coat*
NOTE:
*If the WBC no. of whole blood sample were more than 6 x104
cells/l,
dilute the sample with PBS or saline is recommended
Controls/Optional
internal control#
Add controls /internal control in the extraction procedure if the
downstream analysis needed (# see Controls/internal control on page 18 )
Elute volume 100-400l*
*For those WBCs no. > 30000/l samples, set elute volume more than
200l is recommended
MagPurix Blood DNA Extraction Kit has proven to work
for fresh or frozen blood samples collected in tubes
containing common anti-coagulants like EDTA, heparin*
and citrate(*The EDTA is recommended to use as
anticoagulation agent, while heparin have inhibit effects
on nucleic acid amplification reaction)
If the sample volume is less than 400l or the WBCs no.
is high (more than 60000 cells/l ), add the appropriate
volume of PBS to adjust
Using fresh whole blood sample (within 1 week) for
extraction is recommended, the total nucleic acid yield
would be decreased by time
For short-term storage (up to 10 days), store the tubes
at 2–8°C. However, for applications requiring maximum
fragment size, such as Southern blotting, storage
purified DNA at 2–8°C for up to 3 days , as low levels of
DNA degradation will occur after this time
For long-term storage, store the purified DNA at –70°C.
In fact, the eluate contains total nucleic acid (DNA and
RNA), the RNA is not the major product in this kit (about
10%) and would be degraded soon. If the RNA-free
product is needed, add some RNase to the eluate
www.ZINEXTS.com Page 18 Vr.5.1
Expected Purity and Yield
DNA was purified by using the MagPurix Blood DNA extraction Kit 1200
and the MagPurix Purification Instrument using four different blood samples.
The DNA concentration was measured using a NanoDrop® 2000
spectrophotometer. The range of DNA yield is 2-60g (in the WBCs range 2-60
x 103cells/l)
Sample material Volume/
amount
DNA Yield Purity
Whole Blood
(WBC no. is 6 x 103 /ml)
400l
1000l
9-9.5g
25-30g
OD260/OD280 >=1.7
OD260/OD230 >=1.5
Whole Blood
(WBC no. is 11 x 103 /l)
400l
1000l
18-19g
45-50g
Whole Blood
(WBC no. is 25x 103 /l)
400l
1000l
30-36g
70-75g
Whole Blood
(WBC no. is 40 x 103 /l)
1000l 100g*
Buffy coat 100l
200l
10-30g
20-60g
* For those WBCs count > 30000/l samples, set elute volume more than 200l is
recommended
Controls/ internal control
Using appropriate controls for downstream analysis: :
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control (IC) Using a defined quantity control Place in sample tube or the round
well of the reaction chamber
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Quality Control
In accordance with ZINEXTS’s ISO-certified Quality Management System, each
lot of MagPurix Blood DNA Extraction Kits is tested against predetermined
specifications to ensure consistent product quality
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MagPurix Viral Nucleic Acid Extraction Kit
Cat. No. ZP02003 Process Time: 40-45 minutes
Intended Use
MagPurix Viral Nucleic Acid Extraction Kit is used with the
MagPurix 12 instrument for extraction of Viral DNA or RNA
from human biological specimens such as serum, plasma,
and other cell-free fluids
Application
Nucleic acids extracted from MagPurix Viral Nucleic Acid
Extraction kit can be used in a number of downstream
application including: PCR, qPCR, Sequencing(NGS),
Microarray, RFLP, Southern Blot Analysis
Number Of Tests
36 or 48 extractions
Kit Components
Kit Contents ZP02003-36 ZP02003-48
Reagent Cartridge 36 pcs (6x6) 48 pcs (4x6x2)
Reaction Chamber 36 pcs (6x6) 48 pcs (4x6x2)
Tip Holder 36 pcs 48 pcs (24x2)
Filtered Tip 38 pcs 50 pcs (25x2)
Piercing Pin 38 pcs 50 pcs (25x2)
Sample Tube (2 mL) 38 pcs 50 pcs (25x2)
Elute Tube (1.5 mL) 38 pcs 50 pcs (25x2)
RNA Carrier (1mg) 1 pc 1 pc
Barcode Paper 1 pc 1 pc
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Reagent Cartridge Content
well-1 Proteinase K solution 30 μl
well-2 Lysis Buffer 1 720 μl
well-3 Binding Buffer 1 1000 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 RNase-free water 1000 μl
well-9 Rnase-free water 1000 μl
well-10 Empty
Storage
MagPurix Viral Nucleic Acid Extraction Kit should be
stored at room temperature (15-25°C). Do not freeze
the reagent cartridges. The Kits are stable for 12
months under the condition
After dissolve the carrier RNA, store it at 4°C
(short-term, up to 1 month) or -20°C (long-term). Do
not freeze–thaw the Frozen carrier RNA more than 3
times
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Starting Material
Sample Type Target Nucleic Acid
Sample Volume (Amount of starting material)
Elution Volume
Serum Total Viral
Nucleic Acids
(DNA + RNA)
100-400μl 50-300μl Plasma
CSF
Pretreated Urine
Cell-free body
fluids
Controls/internal
control#
Add controls /internal control in the extraction procedure if
the downstream analysis needed (# see control/internal
control on page 24 )
The kit is designed for extraction of viral nucleic
acids (e.g., those of HIV, HCV, HBV, CMV and
EBV) from plasma or serum, or from a pool of
such cell-free body fluids
After extraction, store the nucleic acid at 4°C
(up to 24hours) or -20°C for longer storage.
Repeated freeze–thawing is not allowed
Sample preparation
The purification procedure is optimized for use
with 100- 400 μl serum, plasma, CSF, or
pretreated urine samples. ( Blood samples
treated with EDTA or citrate as an anticoagulant
can be used for plasma preparation)
Samples can be either fresh or frozen, provided
that they have not been refrozen after thawing
After collection and centrifugation, plasma,
serum, or CSF can be stored at 2–8°C for up to 6
hours. For longer storage, we recommend
freezing aliquots at –20°C or –80°C. Thaw
samples at room temperature (15–25°C), and
www.ZINEXTS.com Page 23 Vr.5.1
process the samples immediately when they
have equilibrated to room temperature. Do not
refreeze the aliquots after thawing. Repeated
freeze–thawing leads to denaturation and
precipitation of proteins, resulting in reduced
viral titers and therefore reduced yields of viral
nucleic acids. If cryoprecipitates are visible in
the samples, centrifuge at 6800 x g for 3
minutes, transfer the supernatants to fresh
tubes without disturbing the pellets, and start
the purification procedure immediately
Carrier RNA For RNA virus, adding carrier RNA to the
sample before extraction is recommended!!!
Add 1.0 ml RNase - free water to the carrier
RNA tube (provided with the kit) and mix by
vortexing. Store it at 4°C (short-term, up to 1
month) or -20°C (long-term). Do not
freeze–thaw the Frozen carrier RNA more than
three times
Add 5 μl Carrier RNA (for 100 μl sample), 10 μl
(for 200 μl sample) or 20 μl (for 400 μl sample)
into the Sample Tube
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Result
(a) DNA virus (HBV)
Using serum spiked with serial-diluted Hepatitis B Virus (in range of
300000-30 IU/ml). 200l sample were extracted and eluted in 100l. 30l
eluate was used for real-time PCR reaction by AmpliSens®
HCV/HBV/HIV-FRT PCR kit. As little of less than 6 IU spiked (about 1 IU in
PCR reaction) sample can be detected, proving the excellent sensitivity
and linearity of isolation procedure
(b) RNA virus (HCV)
Using serum spiked with serial-diluted Hepatitis C Virus (in range of
500000-50 IU/ml). 200l sample were extracted and eluted in 100l. 30l
eluate was used for real-time PCR reaction by AmpliSens®
HCV/HBV/HIV-FRT PCR kit. As little of less than 10 IU spiked (about 3 IU in
PCR reaction) sample can be detected, proving the excellent sensitivity and
linearity of isolation procedure
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Controls/ internal control
Using appropriate controls for downstream analysis:
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control (IC) Using a defined quantity control Place in sample tube or the round well of
the reaction chamber
Quality Control
In accordance with ZINEXTS’s ISO-certified Quality Management System, each lot of
MagPurix Viral Nucleic Acid Extraction Kits is tested against predetermined specifications
to ensure consistent product quality
www.ZINEXTS.com Page 26 Vr.5.1
MagPurix Tissue DNA Extraction Kit
Cat. No. ZP02004 Process Time: 40-45 minutes
Intended Use
MagPurix Tissue DNA Extraction Kit is used with the MagPurix 12 instrument for extraction of genomic DNA from a variety of animal tissues, swab samples and blood stain
For extraction from FFPE samples, using “ZP02009 MagPurix
FFPE DNA extraction kit” is recommended
Application
Nucleic acids extracted from MagPurix Tissue DNA Extraction kit
can be used in a number of downstream application including:
PCR, qPCR, Sequencing(NGS), Microarray, RFLP, Southern Blot
Analysis
Number Of
Tests
36 or 48 extractions
Kit Components
Kit Contents ZP02004-36 ZP02004-48
Reagent Cartridge 36 pcs (6x6) 48 pcs (4x6x2)
Reaction Chamber 36 pcs (6x6) 48 pcs (4x6x2)
Tip Holder 36 pcs 48 pcs (24x2)
Filtered Tip 38 pcs 50 pcs (25x2)
Piercing Pin 38 pcs 50 pcs (25x2)
Sample Tube (2 mL) 38 pcs 50 pcs (25x2)
Elute Tube (1.5 mL) 38 pcs 50 pcs (25x2)
Proteinase K (10mg/mL)
1 pc (1mL) 1 pc (1mL)
BL2 Buffer 1 pc (25mL) 1 pc (25mL)
Barcode Paper 1 pc 1 pc
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Reagent Cartridge Content
well-1 Empty
well-2 Lysis Buffer 2 720 μl
well-3 Binding Buffer 1 720 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 Elution Buffer 1 1000 μl
well-9 Elution Buffer 2 1000 μl
well-10 Empty
Storage
MagPurix Tissue DNA Extraction Kit should be stored at room
temperature (15-25°C). Do not freeze the reagent cartridges.
The Kits are stable for 12 months under the condition
Store the purified DNA at 4 °C (short- term) or aliquot and
store at -70°C (long-term) before perform the downstream
analysis
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Starting
Material
The types and amounts of starting material for use in MagPurix
Tissue DNA purification procedures are shown in Table listed
below,
Sample Type Target
Nucleic Acid
Sample Volume
(Amount of starting material)
Elution
Volume
Tissue DNA 100-400 μl/10-40 mg 50-300μl
Dried swab samples (e.g. Buccal cells)
100-400 μl/1 swab or brush
(add BL2 and proteinase K to
100-400l for extraction)
Dried blood 100-400 μl/4 discs*
Control/Optional
internal control#
Add controls /internal control in the extraction procedure if the
downstream analysis needed (# see control/internal control on page
33 )
*A 3mm diameter disc punched out from filter paper stained with dried
blood contains white blood cells from approximately 5 μl whole blood; we
recommend using 4 punched-out discs as starting material
Yield of
purified DNA
DNA yields depend on the sample type, number of nucleated
cells in the sample, and the protocol used for purification of
DNA
Table listed below shows DNA yields obtained from different
sample types using MagPurix extraction procedures
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Table: The DNA yield of different sample types
Sample Type Sample Amount Typical DNA Yield
Skeletal muscle 200 μl
(40 mg tissue digested)
Up to 9μg
Heart 200 μl
(20 mg tissue digested)
Up to 12μg
Spleen 200 μl
(10 mg tissue digested)
Up to 27μg
Lung 200 μl
(10 mg tissue digested)
Up to 17μg
Kidney 200 μl
(10 mg tissue digested)
Up to 18μg
Liver 200 μl
(10 mg tissue digested)
Up to 40μg
Buccal cells 1 Swab 1-5 μg
Duried blood 4 x 3 mm diameter discs 0.2-0.5 μg
Sample preparation requirements are highly dependent upon the type of
starting material. Due to variations in consistency and viscosity, even similar
sample types may require distinct handling. The steps below describe some
recommendations for processing primary samples
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Sample preparation
For solid animal tissue
���1. �Transfer tissue
No. Sample
Type
Recommended
Sample Amount
1 Heart 20 mg
2 Muscle 40 mg
3 Other
Tissues
10 mg
2. �Add BL2 Buffer
�3. Add Proteinase K
4. �Incubate
5. �Spin down and
Transfer
Spin down and transfer 100-400μl the
supernatant to Sample Tube. Use BL2 buffer to
adjust the sample volume.
OPTIONAL: Use the filter column
(ZA030118, not supplied in kit) to remove
the residual debris and mucous material
before DNA extraction will increase the
DNA yield (20-100%)
Incubate at 55°C in a shaking water bath or
thermomixer (mix set at 1000r.p.m.)until the
tissue is completely lysed.
Note: 1. Lysis time varies depending on the
type of tissue processed. Lysis is usually
completed in 1-2 hrs. However, lysis overnight is
possible and does not influence the preparation.
2. Use the heat block for incubation, vortexing-
mix several times during incubation is
recommend
Transfer tissue into a 1.5ml microcentrifuge
tube:
Add 100-400 μl provided Buffer BL2. Ensure
tissue pieces are fully submerged in Buffer BL2
Add 20 μl proteinase K solution and mix by
vortexing
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For Swap Tissue
�1. Cut����� Carefully cut or break off the end part of
the swab or brush into a 2 ml screw-
capped tube (not supplied) using an
appropriate tool (e.g., scissors).
��������������2. Add BL2 Buffer Add 100-400 μl of Buffer BL2 to the
sample.Ensure sample is fully submerged
in Buffer BL2
�3. Add proteinase K Add 20 μl proteinase K, and mix
thoroughly by vortexing for 10 s.
If processing buccal cell brush samples,
centrifuge the tube briefly (at 10,000 x g
for 30s) to force the brush to the bottom
of the tube.
4. ���Incubate Incubate at 55°C for 15 min (place in a
thermomixer, mix set at 1000 r.p.m. or
votex mix several times during incubation
in heat block)�
�5. Centrifuge Centrifuge the tube briefly to remove
drops from inside the lid.
�6. Remove Remove the swab or brush from the
tube.Using forceps, press the swab or
brush against the inside of the tube to
obtain maximum sample volume. The
sample volume should be approximately
as the setting volume(100-400l). *Use
BL2 buffer to adjust the volume
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For FFPE Tissue
To extract DNA from FFPE samples, please select and refer to
MagPurix FFPE DNA Extraction Kit (ZP02009)
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Dried Blood
Controls/ internal control
Using appropriate controls for downstream analysis: :
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control (IC) Using a defined quantity control Place in sample tube or the round
well of the reaction chamber
1.� Collect Collect 70 μl of each blood sample
onto a ring marked on filter paper.
Allow the blood to air dry.
Either untreated blood or blood
containing anticoagulant (EDTA, ACD,
or heparin) can be used.
2.� Cut For each dried blood sample, use the
manual paper punch to cut out four 3
mm diameter discs.
3.� Add BL2 Transfer each set of 4 discs to a 1.5 ml
microcentrifuge tube. Add 220-440l
BL2 to sample
4.� Add
proteinase K
Add 20l proteinase K and mix by
vortexing
5.� Incubation incubate at 55℃, 15min in a
thermomixer (set mix at 1000r.p.m.)
or vortex mix several time during
incubation in heat block
6.� Centrifuge Centrifuge the tube brifly to remove
the drop inside the lid
7.� Transfer Transfer the 100-400l supernatant to
the sample tube (provided in the kit),
proceeding DNA extraction
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Quality Control
In accordance with ZINEXTS’s ISO-certified Quality Management System, each lot of
MagPurix Tissue DNA Extraction Kit is tested against predetermined specifications to
ensure consistent product quality.
www.ZINEXTS.com Page 35 Vr.5.1
MagPurix Cultured Cell DNA Extraction Kit
Cat. No. ZP02005 Process Time: 40- 45 minutes
Intended Use
MagPurix Cultured Cell DNA Extraction Kit is used with the
MagPurix 12 instrument for extraction of genomic DNA
from culture cells and buffy coat
Application
Nucleic acids extracted from Cultured Cell DNA Extraction
kit can be used in a number of downstream application
including: PCR, qPCR, Sequencing(NGS), Microarray, RFLP,
Southern Blot Analysis
Number Of
Tests
36 or 48 extractions
Kit
Components
Kit Contents ZP02005-36 ZP02005-48
Reagent Cartridge 36 pcs (6x6) 48 pcs (4x6x2)
Reaction Chamber 36 pcs (6x6) 48 pcs (4x6x2)
Tip Holder 36 pcs 48 pcs(24x2)
Filtered Tip 38 pcs 50 pcs(25x2)
Piercing Pin 38 pcs 50 pcs(25x2)
Sample Tube (2 mL) 38 pcs 50 pcs(25x2)
Elute Tube (1.5 mL) 38 pcs 50 pcs(25x2)
Barcode Paper 1 pc 1 pc
www.ZINEXTS.com Page 36 Vr.5.1
Reagent Cartridge Content
well-1 Proteinase K solution 40 μl
well-2 Lysis Buffer 2 720 μl
well-3 Binding Buffer 1 720 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 Elution Buffer 1 1000 μl
well-9 Elution Buffer 2 1000 μl
well-10 Empty
Storage
MagPurix Cultured Cell DNA Extraction Kit should be
stored at room temperature (15-25 °C). Do not freeze
the reagent cartridges. The Kits are stable for 12
months under the condition
Store the purified DNA at 4 °C (short- term) or aliquot
and store at -70°C (long-term) before perform the
downstream analysis
www.ZINEXTS.com Page 37 Vr.5.1
Starting
Material
1. Culture cells in suspension and monolayer
2. Cells from Buffy coat (without red blood cells)
If the buffy coat is remove directly from the whold
blood, for completely remove and lyse red blood cell,
the MagPurix Blood DNA Extraction kit (ZP02001;
ZP02002) is recommended
Do not use more than 5 x 106 cells with a normal set
of chromosomes
The cell number can be determined by using
Hemocytometer1,2 (Petroff-Hauser Chamber )and
automated cell counter( e.g. TC10™, Countess®,
Cellometer®, and Scepter™ automated cell
counters )
The types and amounts of starting material for use in
MagPurix Cultured Cell DNA purification procedures
are shown in Table listed below
Reference
1. http://www.smccd.edu/accounts/case/biol230/algae/hemo
cytometer1.pdf
2. http://web.mnstate.edu/provost/CountingCellsHemocytom
eter.pdf
Sample preparation
Cells grown in suspension
Suspension culture:
1. Determine the number of cells. ( Do not use
more than 5 x 106 cells with a normal set of
chromosomes)
2. Centrifuge the appropriate number of cells for 5
min at 1000 x g in a 1.5 ml microcentrifuge tube
3. Remove the supernatant completely and discard,
taking care not to disturb the cell pellet
4. Resuspend cell pellet in PBS to a final volume of
200 μl
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Cells grown in a monolayer
Cells grown in a monolayer can be detached from
the culture flask by either trypsinization or using a
cell scraper
To trypsinize cells:
1. Determine the number of cells* (do not use more
than 5 x 106 cells with a normal set of
chromosomes)
2. Aspirate the medium and wash cells with PBS
3. Aspirate the PBS, and add 0.10–0.25% trypsin,
incubate at 37°C
4. After cells have detached from the dish or flask,
collect them in medium, and transfer the
appropriate number of cells to a 1.5 ml
microcentrifuge tube.
5. Centrifuge for 5 min at 1000 x g.
6. Remove the supernatant completely and discard,
taking care not to disturb the cell pellet.
7. Resuspend cell pellet in PBS to a final volume of
200 μl.
Using a cell scraper:
1. Determine the number of cells. (do not use more
than 5 x 106 cells with a normal set of
chromosomes)
2. Detach cells from the dish or flask by scraping
3. Harvest and transfer cells to a 1.5 ml
microcentrifuge tube and centrifuge for 5 min at
1000 x g.
4. Remove the supernatant completely and discard,
taking care not to disturb the cell pellet.
5. Resuspend cell pellet in PBS to a final volume of
200 μl
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Yield Of
Purified DNA
DNA yields depend on the sample type, number of
nucleated cells in the sample, and the protocol used for
purification of DNA.
For example, the average DNA yield form the HT29 colon
adenocarcinoma cell line at the different concentrations
( in the range from 1 x 105 to 106 cells) is about 22g/106
cells as below
Quality Control
In accordance with ZINEXTS’s ISO-certified Quality Management System,
each lot of MagPurix Cultured Cell DNA Extraction Kits is tested against
predetermined specifications to ensure consistent product quality
www.ZINEXTS.com Page 40 Vr.5.1
MagPurix Bacterial DNA Extraction Kit
Cat. No. ZP02006 Process Time: 55-65 minutes
Intended Use
MagPurix Bacterial DNA Extraction Kit is used with the MagPurix 12 instrument for extraction of genomic DNA from both Gram-positive and Gram-negative bacteria
Application
Nucleic acids extracted from Bacterial DNA Extraction kit can be used in a number of downstream application including: PCR, qPCR, Sequencing(NGS), Microarray, RFLP, Southern Blot Analysis
Number Of
Tests
36 or 48 extractions
Kit
Components
Kit Contents ZP02006-36 ZP02006-48
Reagent Cartridge 36 pcs (6x6) 48 pcs (4x6x2)
Reaction Chamber 36 pcs (6x6) 48 pcs (4x6x2)
Tip Holder 36 pcs 48 pcs (24x2)
Filtered Tip 38 pcs 50 pcs (25x2)
Piercing Pin 38 pcs 50 pcs (25x2)
Sample Tube (2 mL) 38 pcs 50 pcs (25x2)
Elute Tube (1.5 mL) 38 pcs 50 pcs (25x2)
BL2 Buffer 1 pc (25mL) 1 pcs (25mL)
Barcode Paper 1 pc 1 pc
www.ZINEXTS.com Page 41 Vr.5.1
Reagent Cartridge Content
well-1 Proteinase K solution 40 μl
well-2 Lysis Buffer 2 720 μl
well-3 Binding Buffer 1 720 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 Elution Buffer 1 1000 μl
well-9 Elution Buffer 2 1000 μl
well-10 Empty
Storage
MagPurix Bacterial DNA Extraction Kit should be stored at
room temperature (15-25°C). Do not freeze the reagent
cartridges.The Kits are stable for 12 months under the
condition
Store the purified DNA at 4 °C (short- term) or aliquot and
store at -70°C (long-term) before perform the downstream
analysis
www.ZINEXTS.com Page 42 Vr.5.1
Starting
Material
Bacterial pellet/colony from culture, cell-free body fluids,
liquid transport media, urine, environment material (water, soil, etc.)
Use the paraffin- embedded tissue sections as samples, we recommend to extract DNA by MagPurix FFPE DNA Extraction kit (ZP02009)
Use tissue as samples, we recommended use the MagPurix Tissue DNA Extraction kit
The types and amounts of starting material for use in
MagPurix Bacterial DNA purification procedures are shown in Table listed below,
Sample Type Target Nucleic Acid
Sample volume (Amount of starting material)
Elution Volume
Bacteria Pellet Genomic DNA 200-400 μl /Up to 109
bacteria (about OD600 = 3)
50-300 μl
Bacterial colony 200-400 μl /1-3 colony
Tissue 200-400 μl /1-30 mg
Urine 200-400 μl /5-50 mL urine
Cell-free body fluids
200-400 μl cell-free body fluids
Liquid transport media
200-400 μl liquid transport media
NOTE: Before extraction, adjust sample volume with BL2 buffer
Sample
Preparation
Sample preparation requirements are highly dependent
upon the type of starting material. Due to variations in consistency and viscosity, even similar sample types may require distinct handling
The buffer BL2 is specialized for bacterial cell wall lyse* (Supplied in the kit), use it to resuspend the bacterial pellet before process extraction
* For mycobacterium spp.(e.g. MTB), use buffer BL3 for
www.ZINEXTS.com Page 43 Vr.5.1
Table : Preparation of sample material for bacterial nucleic acid extraction
Sample type Procedure
For viscous samples e.g. BAL, sputum or other mucous specimen
Recommended pretreatment : Liquefaction 1. Prepare a fresh DTT stock solution for
liquefaction * (e.g., 5× conc. DTT stock is about 0.75%)
2. Adjust the final DTT concentration in the sample
to 0.15% by adding DTT stock solution.
3. Incubate the sample (e.g., with shaking at 850r.p.m. for 30 min at 37°C) until it can be pipette easily.
4. Pellet bacteria by centrifugation at 14000 x g for 10 min
5. Discard supernatant, resuspend the pellet in 220 μL Buffer BL2
6. Transfer 200 μL suspension to sample tube (Supplied in the kit) * The liquefaction could be done by using other solutions, such as NALC(N-Acetyl-L-Cysteine)
-NaOH or other agents which could digest mucous material
bacterial cell wall lysis (BL3 buffer is supplied in the ZP02008
MagPurix TB DNA Extraction kit )
The table below describes the recommendations in processing the primary samples before nucleic acid extraction:
www.ZINEXTS.com Page 44 Vr.5.1
For large volume liquid samples that have low or unknown bacterial loads e.g. urine, water collected from pool/river stream/tower
Recommended pretreatment : Centrifugation 1. Centrifuge the sample for up to 10 min at
20,000 × g to concentrate the bacterial cells in pellet
2. Discard supernatant, resuspend the pellet in 220 μL Buffer BL2*
3. Take 200 μL suspension to sample tube (Supplied in the kit)
* If there were sand or other visible particle in the pellet, centrifuge again after BL2 buffer treatment or filter out the dust is recommended
For cell-free body fluids (e.g. CSF, BAL, aspirates)
Recommended pretreatment : Centrifugation Method 1 1. Pellet bacteria by centrifugation at 14000 x g
for 10 min 2. Resuspend bacterial pellet in 220 μL Buffer BL2 3. Take 200 μL suspension to sample tube
(Supplied in the kit) Method 2-Centrifugation free
1. Take 200 l sample in a 1.5 ml centrifuge tube
2. Add 200l buffer BL2 to sample (1:1) 3. Vortex-mixing for 5-10sec
4. Transfer 400l sample to sample tube (Supplied in the kit)
For swab samples e.g. eye, nasal, pharyngeal, or other swabs
Method 1 1. Collect samples and place in 2 ml PBS
containing a common fungicide. Incubate for 30min at room temperature
2. Pellet bacteria by centrifugation at 14000 x g for 10 min
3. Resuspend bacterial pellet in 220 μL Buffer BL2
(Supplied in the kit)
4. Take 200 μL suspension to sample tube (Supplied in the kit)
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Method 2- centrifuge free
1. Place the sample swab in 440l buffer BL2, incubate for 30min at room temperature
2. Transfer 400l to sample tube
For some gram-positive bacteria species. Especially for samples that contain particles e.g. stool
Recommended pretreatment : Mechanical homogenization
Follow the regular homogenization procedures
in the laboratory.
For some sample types, DNA yield can be
improved by performing this homogenization
step prior to add buffer BL2 and proteinase K
Isolation of genomic DNA from bacterial suspension cultures
1. Pipet 1 ml of bacterial culture into a 1.5 ml
microcentrifuge tube and centrifuge at 5000xg for 5 min
2. Discard supernatant
3. Add 220l Buffer BL2 to pellet and mix by vortexing for 5-10 sec
4. Take 200 μL suspension to sample tube
(Supplied in the kit)
Isolation of genomic DNA
from bacterial plate
culture
1. Take 1-3 bacterial colony from culture plate
with an inoculation loop and suspend in 220 μL of buffer BL2 by vigorous stirring
2. Take 200 μL suspension to sample tube (Supplied in the kit)
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To inactive pathogenic
organisms in the sample
Recommended pretreatment : Boiling
1. Incubate samples at 95°C for 10 min
2. Centrifuge briefly to collect the complete
sample volume at the bottom of the tube.
3. Allow samples to cool down or chill on ice,
then transfer 100-400l cooled sample to the
sample tube
Yield of purified DNA
DNA yields depend on the sample type, number of bacteria in the sample, and the protocol used for purification of DNA
Result
(1) Scalability Using MagPurix Bacterial DNA Extraction kit to isolate the DNA from
cultured Escherichia. coli (ATCC25922) and Staphylococcus aureus
(ATCC27154) in LB broth at different bacterial density (measure the Optical
Density at 600nm; OD600). Take 200l bacterial culture for extraction and
collect the eluate in 100l. The total nucleic acid yield of different bacterial
density was measured by Nanodrop 2000 UV-Vis spectrophotometer (fig.1a
and 2a) and analyzed by 1% TAE agarose gel electrophoresis (fig.1b and 2b).
The result shown the nucleic acid extraction in both Gram-negative (E.coli)
and Gram-positive (S. aureus) were have excellent scalability.
0
5
10
15
20
0 0.5 1 1.5 2 2.5
tota
l nu
cle
ic a
cid
yie
ld (
g)
Turbidity (O.D.600)
Fig.1a E. coli
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Fig.1b
Fig.2b
(2) Sensitivity Performing serial-dilution on Staphylococcus aureus (ATCC27154) in range
of 109-101copy/ml). 200l sample were extracted and eluted in 100l. 25l
eluate was used for SYBR Green real-time PCR reaction which detect
Staphylococcus aureus specific gene. As little of 20 copies (about 102
copy/ml bacteria in the sample) spiked-in (about 5 copy in PCR reaction)
bacteria can be detected, proving the excellent sensitivity and linearity of
isolation procedure (fig.3a and 3b)
0
5
10
15
0 0.5 1 1.5 2 2.5 3
tota
l nu
cle
ic a
cid
yie
ld (
g)
Turbidity (O.D.600)
Fig.2a Staphylococcus aureus
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Fig. 3a
Fig.3b
Controls/ internal control
Using appropriate controls for downstream analysis:
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control (IC) Using a defined quantity control Place in sample tube or the round
well of the reaction chamber
Quality Control In accordance with ZINEXTS’s ISO-certified Quality Management System,
each lot of MagPurix Bacterial DNA Extraction Kits is tested against
predetermined specifications to ensure consistent product quality
www.ZINEXTS.com Page 49 Vr.5.1
MagPurix HPV DNA Extraction Kit for
Swab samples
Cat. No. ZP02007 Process Time: 40-50minutes
Intended Use
MagPurix HPV DNA Extraction Kit is used with the MagPurix 12
instrument for DNA extraction of the Human Papillomavirus
(HPV) from cervical cell samples which collected by cervical
brush or genital swab in liquid-based Medium* (e.g. Hologic
Thinprep PreservCyt®, BD SurepathTM, etc.) or other STM
(sample transport media) preservation solutions(e.g. QIAGEN
DNA PAP Cervical sampler, Roche Cobas® PCR Cell Collection
Media, Hybribio cell preservation solution, etc.).
*The liquid-base medium are formulated for cellular
preservation and used in liquid-based cytological systems(LBC)
for cytological and molecular diagnosis
Application
Nucleic acids extracted from HPV DNA Extraction kit from swab sample can be used in a number of downstream application including: PCR, qPCR, Sequencing (NGS), Microarray, RFLP, Southern Blot Analysis, etc.
Number Of
Tests
36 or 48 extractions
www.ZINEXTS.com Page 50 Vr.5.1
Reagent Cartridge Content
well-1 Proteinase K solution 40 μl
well-2 Lysis Buffer 2 720 μl
well-3 Binding Buffer 1 1000 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 Elution Buffer 1 1000 μl
well-9 Elution Buffer 2 1000 μl
well-10 Empty
Kit
Components
Kit Contents ZP02007-36 ZP02007-48
Reagent Cartridge 36 pcs (6x6) 48 pcs (4x6x2)
Reaction Chamber 36 pcs (6x6) 48 pcs (4x6x2)
Tip Holder 36 pcs 48 pcs (24x2)
Filtered Tip 38 pcs 50 pcs (25x2)
Piercing Pin 38 pcs 50 pcs (25x2)
Sample Tube (2 mL) 38 pcs 50 pcs (25x2)
Elute Tube (1.5 mL) 38 pcs 50 pcs (25x2)
BL4 Buffer 1 pc (25mL) 1 pcs (25mL)
Barcode Paper 1 pc 1 pc
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Storage
MagPurix HPV DNA Extraction Kit for swab
samples should be stored at room temperature
(15-25°C). Do not freeze the reagent cartridges.
The Kits are stable for 12 months under the
condition
Store the purified DNA at 4 °C (short- term) or
aliquot and store at -70°C (long-term) before
perform the downstream analysis
Starting Material
Cervical cells collected by cervical brush or
genital swab
For those non-liquid based medium, adding
BL4 Buffer to the reservation is
recommended
The specimen should be sent at 4-30°C for
examination on immediately after collection.
The storage condition is depended on the
preservation solution
Add controls /internal control in the
extraction procedure if the downstream
analysis needed (see control/internal control
on page 52 )
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Sample Preparation
Sample type Procedure
In liquid-based
preservation solution
(eg. Hologic Thinprep
PreservCyt®, BD
SurepathTM)
1. Take the sample amount as the following assay
recommended*
2. Centrifuge at 1000 x g. for 5min
3. Discard supernatant
4. Resuspend pellet in 220l BL4
5. Incubate at RT, 5 min
6. Vortex for 5 sec
7. Take 200 l suspension for extraction
In other STM
preservation solution
(QIAGEN DNA PAP, Hybribio
cell preservation solution)
1. Add equal volume of BL4 directly to the reservation
solution (BL4: reservation = 1:1) **
2. Incubate at RT, 5-10 min
3. Vortex for 5 sec
4. Take 100-400 l sample for extraction
* Especially for those assay of” signal amplification”, in that the target DNA won’t be
amplified. Take adequate amount of sample is necessary
** The sticky mucus is common in cervical specimen, adding BL4 before processing
will help sample liquefying and nucleic acid extraction
Controls/ internal control
Using appropriate controls for downstream analysis: :
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control (IC) Using a defined quantity control Place in sample tube or
the round well of the
reaction chamber
www.ZINEXTS.com Page 53 Vr.5.1
Quality Control
In accordance with ZINEXTS’s ISO-certified Quality Management System, each lot of
MagPurix HPV DNA Extraction Kits for swab samples is tested against predetermined
specifications to ensure consistent product quality
www.ZINEXTS.com Page 54 Vr.5.1
MagPurix TB DNA Extraction Kit
Cat. No. ZP02008 Process Time: 60-70 minutes
Intended use
MagPurix TB DNA Extraction Kit is used with the MagPurix 12
instrument for extraction of genomic DNA of Mycobaceteria
spp. (e.g. Mycobacterium tuberculosis) from different specimen
Application
Nucleic acids extracted from TB DNA Extraction kit can be used
in a number of downstream application including: PCR, qPCR,
Sequencing(NGS), Microarray, RFLP, Southern Blot Analysis
Number Of
Tests
36 or 48 extractions
Kit
Components
Kit Contents ZP02008-36 ZP02008-48
Reagent Cartridge 36 pcs (6x6) 48 pcs (4x6x2)
Reaction Chamber 36 pcs (6x6) 48 pcs (4x6x2)
Tip Holder 36 pcs 48 pcs (24x2)
Filtered Tip 38 pcs 50 pcs (25x2)
Piercing Pin 38 pcs 50 pcs (25x2)
Sample Tube (2 mL) 38 pcs 50 pcs (25x2)
Elute Tube (1.5 mL) 38 pcs 50 pcs (25x2)
BL3 Buffer 1 pc (25mL) 1 pc (25mL)
Barcode Paper 1 pc 1 pc
www.ZINEXTS.com Page 55 Vr.5.1
Reagent Cartridge Content
well-1 Proteinase K solution 40 μl
well-2 Lysis Buffer 2 720 μl
well-3 Binding Buffer 1 720 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 Elution Buffer 1 1000 μl
well-9 Elution Buffer 2 1000 μl
well-10 Buffer N1 400 μl
Storage
MagPurix TB DNA Extraction Kit should be stored
at room temperature (15-25 °C). Do not freeze
the reagent cartridges. The Kits are stable for 12
months under the condition
Store the purified DNA at 4 °C (short- term) or
aliquot and store at -70°C (long-term) before
perform the downstream analysis
Starting Material
1. Clinical specimen: Sputum, BAL, Pas, blood,
cell-free body fluids, urine and other respiratory
specimens
2. Bacterial culture in the solid and liquid media For the MTB is highly infectious agent, prepare
sample in the BSC(Biosafety cabinet)
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Sample preparation
Specimen type
Procedure
Sputum/BAL or other
Respiratory Specimen
Method 1
1. Liquify the sample*
2. Pellet bacteria by centrifugation at 12500 x g
for 15 min
3. Discard supernatant, resuspend the pellet in
200 μL Buffer BL3, votex mixing about 5 sec
4. Take 200 l sample to sample tube for
extraction
Method 2 - Centrifuge free
1. Liquify the sample*
2. Transfer 200l sample to sample tube
3. Add 200l buffer BL3 to sample (1:1)
1.
* The liquefaction could be done by using
liquefying agents, such as NALC(N-Acetyl-L-
Cysteine)- NaOH and 0.75% DTT (5x stock)
which could digest mucous material
Viscous body fluid e.g. Pas
See the procedure of “Sputum/BAL or other
Respiratory Specimen”
Cell-free body fluid
e.g. CSF, urine
1. Pellet bacteria by centrifugation at 14000 x g
for 15 min
2. Discard supernatant, resuspend bacterial
pellet in 200 μL Buffer BL3,Votex-mixing
about 5 sec
3. Take 200 μL sample to sample tube for
extraction
Liquefied,
decontaminated sample
See the procedure of “cell-free body fluid”
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Blood or
Blood-contaminated
sample
1.
1. Add cold sterilized water to sample to the
ratio of water/blood about 3:1
2. Inverted mix several times
3. Incubate at 4℃, at least 10 min
4. Centrifuge at 14000 x g for 15 min
5. Remove supernatant, add 200 μL buffer BL3,
votex mixing about 5-10sec
6. Take 200 μL sample to sample tube for
extraction
Colony from solid
culture
1. Pick 1-3 colony, mix with 200 μL Buffer BL3,
votex mixing about 5-10 sec
2. Take 200 μL sample to sample tube for
extraction
Liquid culture Method 1
1. Take 1mL culture(>McFarland 0.5), transfer
to 1.5mL microcentrifuge tube
2. Pellet bacteria by centrifugation at 12500 x g
for 5 min
3. Discard supernatant, resuspend bacterial
pellet in 200 μL Buffer BL3,Votex mixing
about 5-10 sec
4. Take 200 μL sample to sample tube for
extraction
Method 2- centrifuge free
1. Add Buffer BL3 to liquid culture(1:1)
2. Votex mixing about 5-10 sec
3. Take sample mixture to sample tube for
extraction
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Controls/ internal control
Using appropriate controls for downstream analysis:
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control (IC) Using a defined quantity control Place in sample tube or the round
well of the reaction chamber
Result
Using MagPurix TB DNA Extraction kit to isolate DNA from clinical specimens
(sputum, CSF and pas). 100ml sample used for extraction and collect 100ml eluate.
Analysis was performed by real-time qPCR with Taqman probe /primers (IS6110).
Even in the cell-free body fluid (CSF) and blood contaminated sample(Pas), the TB
DNA can be detected after extraction, proving the excellent sensitivity of isolation
procedure
Expected Purity and Yield
DNA yields depend on the sample type, number of bacteria in the sample, and the
protocol used for purification of DNA
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Quality Control
In accordance with ZINEXTS’s ISO-certified Quality Management System,
each lot of
MagPurix TB DNA Extraction Kits is tested against predetermined
specifications to ensure consistent product quality
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MagPurix FFPE DNA Extraction Kit
Cat. No. ZP02009 Process Time: 35-45 minutes
Intended
Use
MagPurix FFPE DNA Extraction Kit is used with the MagPurix 12 instrument for extraction of genomic DNA from FFPE (Formalin-Fixed, Paraffin-Embedded) tissue samples. Providing good quality, high integrity DNA for Molecular diagnosis and research works
Application
Nucleic acids extracted from FFPE DNA Extraction kit can be used in a
number of downstream application including: PCR, qPCR,
Sequencing(NGS), Microarray, RFLP, Southern Blot Analysis
Number Of
Tests
36 or 48 extractions
Kit
Components
Kit Contents ZP02009-36 ZP02009-48
Reagent Cartridge 36 pcs (6x6) 48 pcs (4x6x2)
Reaction Chamber 36 pcs (6x6) 48 pcs (4x6x2)
Tip Holder 36 pcs 48 pcs (24x2)
Filtered Tip 38 pcs 50 pcs (25x2)
Piercing Pin 38 pcs 50 pcs (25x2)
Sample Tube (2 mL) 38 pcs 50 pcs (25x2)
Elute Tube (1.5 mL) 38 pcs 50 pcs (25x2)
Proteinase K (10mg/mL) 1 pc (1mL) 2 pcs (0.6mL,1x2)
BL2 Buffer 1pc (25 mL) 1 pcs (25mL)
Filter Column 38 pcs 50 pcs (25x2)
Collection Tube 38 pcs 50 pcs (25x2)
Barcode Paper 1 pc 1 pc
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Reagent Cartridge Content
well-1 Empty
well-2 Lysis Buffer 2 720 μl
well-3 Binding Buffer 1 720 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 Elution Buffer 1 1000 μl
well-9 Elution Buffer 2 1000 μl
well-10 Empty
Storage
MagPurix FFPE DNA Extraction Kit should be stored at room
temperature (15-25 °C). Do not freeze the reagent cartridges. The Kits
are stable for 12 months under the condition
Store the purified DNA at 4 °C (short- term) or aliquot and store at
-70°C (long-term) before perform the downstream analysis
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Starting
Material
FFPE (formalin fixed Paraffin Embedded) tissue samples: one to five
10m-thick section(s)
Sample Type Target
Nucleic Acid
Sample Volume
(Amount of starting material)
Elution
Volume
FFPE (formalin fixed Paraffin Embedded) tissue samples
DNA 100-400μl/ One to eight 10 μm-
thick sections (after proteinase
K digestion)*
50-300l
Needle biopsy 100-400μl/ three to ten
biopsies
*Note: The size of tissue area should be more than 1 x 1 cm2, if the area is smaller, use
more than one section for extraction.
Yield of
purified
DNA
DNA yields depend on the sample type, number of nucleated cells in
the sample, and the thickness of the section
Sample preparation requirements are highly dependent upon the
type of starting material. Due to variations in consistency and
viscosity, even similar sample types may require distinct handling.
The steps below describe some recommendations for processing
primary samples
The DNA of FFPE tissue sample is often fragmented which cause
problems in molecular assay. Keep the integrity of DNA is most
important in whole procedure.
Because of the DNA fragments often cause high OD260 and ratio of
260/280. The integrity of FFPE DNA can’t be determined only by
UV-VIS spectrophotometer analysis. The best method for the
integrity check is performing a PCR reaction of house-keeping genes
with different length products
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Sample preparation
1. Transfer sections Transfer sections of paraffin-embedded tissue into a 1.5ml
microcentrifuge tube
*for most tissue types, slice of one to five 10mm-thick sections
is recommened; however, up to ten very small sections such as
needle biopsies may be used
2. Deparaffin
(Optional)
Treat FFPE samples with xylene or other deparaffin agents
*This step is optional, skip this step is still work for the
extraction
3. Add BL2 Buffer Add 400l BL2, ensure that the samples are fully submerged in
BL2 buffer
4. Add proteinase K Add 20ml Proteinase K to sample mixture, vortex mixing for
5-10 sec. short spin for remove the drop from the lid
5. Incubation Incubate sample at 55℃ for 2 hours, with vigorous mixing in a
shaking water bath or thermomixer
*The first 2 hour is the critical step for proteinase lysis.
Incubating sample with shaker is necessary (or vortex mixing
every 30 minutes) for completely digestion. Increase the
incubation time is not much helpful for lysis, because the
proteinase activity drops after 2 hours. Only when the tissue
sample is thick, add more proteinase K and incubate longer is
recommended
6. Centrifuge Centrifuge the tube briefly to remove drops from inside the lid
7. Homogenize Homogenize the sample by pipetting up and down several
times. Transfer the sample to the filter column sitting in a
collection tube (supplied in the kit). Centrifuge at 6000xg, 1min
*Pieces of insoluble material, which could clog tips, make
interferences of extraction efficiency. Remove them by filter
8. Transfer Transfer 400l sample to sample tube
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Result
Integrity
Take eleven Myeloma FFPE section samples (10m thick, about 2 x 2 cm2) for extraction.
Two sections were extracted by QIAGEN FFPE Tissue kit; the others were extracted by
MagPurix FFPE DNA Extraction kit. Using DNA ladder PCR (a multiplex primer PCR of
different length products of house-keeping gene) and capillary electrophoresis to analyze
the integrity of the extracts of PCR products(fig.1)
Fig. 1 The results of capillary electrophoresis shown a quirt good integrity of the extract
Fig.1a QH and QA were the extracts from QIAGEN FFPE tissue kit (elute in water and ATE)
and extracts from MagPurix FFPE DNA were labeled as no. 1 to 9
Scalability
Using Myeloma FFPE section samples (10m thick, about 2 x 2 cm2) for extraction.
Measuring the eluate by UV-VIS Nanodrop2000 spectrophotometer
No. of section Con. (ng/l) Spectrum
1/4 section 1.5
1/2 section 3.8
1 section 8.3
2 section 15.8
3 section 24.7
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Controls/ internal control
Using appropriate controls for downstream analysis:
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control (IC) Using a defined quantity control Place in sample tube or the round
well of the reaction chamber
Quality Control In accordance with ZINEXTS’s ISO-certified Quality Management System, each lot of
MagPurix FFPE DNA Extraction Kits is tested against predetermined specifications to
ensure consistent product quality
www.ZINEXTS.com Page 66 Vr.5.1
MagPurix Forensic DNA Extraction Kit
Cat. No. ZP02010 Process Time: 40-50 minutes
Intended
Use
MagPurix Forensic DNA extraction kit is extract and isolate
genomic DNA from forensic samples
Application
The extracted DNA is compatible for use in quantitation using
the Quantifiler® Human, Quantifiler® Y Human Male, Quantifiler®
Duo DNA Quantification Kits and Investigator® Quantiplex kit, and
for use in STR amplification using the AmpFlSTR® PCR
Amplification kits
Number Of
Tests
36 or 48 extractions
Kit
Components
Kit Contents ZP02010-36 ZP02010-48
Reagent Cartridge 36 pcs (6x6) 48 pcs (4x6x2)
Reaction Chamber 36 pcs (6x6) 48 pcs (4x6x2)
Tip Holder 36 pcs 48 pcs (24x2)
Filtered Tip 38 pcs 50 pcs (25x2)
Piercing Pin 38 pcs 50 pcs (25x2)
Sample Tube (2 mL) 38 pcs 50 pcs (25x2)
Elute Tube (1.5 mL) 38 pcs 50 pcs (25x2)
Proteinase K (10mg/mL)
1 pc (1mL) 2 pcs (0.6mL,1x2)
Buffer BL2 1pc (25 mL) 1 pc (25mL)
Filter Column 38 pcs 50 pcs (25x2)
Collection Tube 38 pcs 50 pcs (25x2)
Barcode Paper 1 pc 1 pc
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Reagent Cartridge Content
well-1 Empty
well-2 Lysis Buffer 1 1000 μl
well-3 Binding Buffer 1 1000 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 Elution Buffer 1 1000 μl
well-9 Elution Buffer 2 1000 μl
well-10 Empty
Storage
MagPurix Forensic DNA Extraction Kit should be
stored at room temperature (15-25 °C). Do not
freeze the reagent cartridges. The Kits are stable
for 12 months under the condition
Store the purified DNA at 4 °C (short- term) or
aliquot and store at -70°C (long-term) before
perform the downstream analysis
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Starting Material
Whole blood, clotted/dried blood, Forensic Surface and Contact Swabs, hair roots,
saliva, sperm stain, chewing gum, cigarette butts, stamps, envelopes, tissue etc.
Sample preparation
Sample type Procedure
Whole blood
(fresh or frozen)
To extract DNA from whole blood samples, please select and refer to MagPurix Blood DNA Extraction Kit 200 (ZP02001)
Clotted/ dried blood 1. Take 20μl blood sample to filter paper or bandage
2. Air-drying the blood sample
3. Cut the blood-contain range out, transfer pieces to
sample tube
4. Add 400μl BL2 and 20μl proteinase K to sample
5. Incubate at 56°C for 15min, vortex-mixing several
times during incubation or place sample in a
thermomixer
6. Transfer all the sample to filter column sitting in a
sample tube
7. Short spin at 500 x g, 1min
8. Take the sample tube for extraction
Forensic Surface and
Contact Swabs
1. Allow the swab or brush to air-dry for at least 2
hours after collection
2. Carefully cut or break off the end part of the swab
or brush into a 1.5ml microcentrifuge tube, using
an appropriate tool (e.g., scissors)
3. Add 200 or 400 μl of Buffer BL2 to the sample
4. Add 20 μl proteinase K, vortex-mixing for at least
10s
*If processing brush samples, centrifuge the tube
briefly (at 10,000 x g for 30 s) to force the brush to the
bottom of the tube
5. Incubate at 56°C for 15 min. vortex-mixing several
times during incubation or place sample in a
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thermomixer
6. Transfer all the sample to filter column sitting in a
sample tube
7. Short spin at 500 xg, 1min
8. Take the sample tube for extraction
Hair root Using two or three 0.5–1 cm from the root ends of
plucked hair samples
【method1】
1. Place the hair sample in a 1.5 ml microcentrifuge
tube
2. Add 200μl Buffer BL2 to the sample.
3. Add 20μl proteinase K and 10 μl 1M DTT solution*,
and mix thoroughly by vortexing for at least 10 s
4. Incubate at 56°C for at least 6 h, vortex-mixing
several times during incubation or place sample in
a thermomixer
5. (optional) Add extra 10 μl proteinase K and 10 μl
DTT and incubate at 56°C until the hair samples are
completely dissolved
6. Spin the tube to remove drops from inside the lid
7. Transfer all the sample to filter column sitting in a
sample tube
8. Short spin at 500 xg, 1min
9. Take the sample tube for extraction
*Prepare 1M DTT solution before processing the
protocol (1M is about 15% DTT(m/v))
【method2】
1. Place the hair sample in a 1.5 ml microcentrifuge
tube
2. Add 200 μl Buffer BL2 to the sample.
3. Add 20 μl proteinase K ,mix thoroughly by
vortexing for at least 10 s
4. Incubate at 56°C overnight, vortex-mixing several
times during incubation or place sample in a
thermomixer
5. Spin the tube to remove drops from inside the lid
6. Transfer all the sample to filter column sitting in a
sample tube
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7. Short spin at 500 xg, 1min
8. Take the sample tube for extraction
Human tissues Using up to 40mg tissue
1. Place tissue sample into a 1.5 ml microcentrifuge
tube
2. Add 200 or 400μl buffer BL2 and 20μl proteinaseK
to the sample, mix thoroughtly by vortexing for 10s
3. Incubate at 56°C for at least 2 hours,*
vortex-mixing several times during incubation or
place sample in a thermomixer
*Incubation for longer time (e.g. overnight) isn’t
making interferences of nucleic acid extraction
4. Spin the tube to remove drops from inside the lid
5. Transfer all the sample to filter column sitting in a
sample tube
6. Short spin at 500 xg, 1min
7. Take the sample tube for extraction
Saliva 1. Place up to 50 μl saliva in a 1.5ml microcentrifuge
tube
2. Add 200 μl Buffer BL2 to the sample
3. Add 20 μl proteinase K, and mix thoroughly by
vortexing for 10 s
4. Incubate at 56°C for 15min, vortex-mixing several
times during incubation or place sample in a
thermomixer
5. Spin the tube to remove drops from inside the lid
6. Take 200μl to sample tube for extraction
Sperm stains 1. Place 5-10μl or 1cm2 of the forensic sample in a 1.5
ml centrifuge tube
2. Add 200 or 400μl Buffer BL2 to the sample
3. Add 20 μl proteinase K, and mix thoroughly by
vortexing for 10 s
4. Incubate at 56°C for 15 min, vortex-mixing several
times during incubation or place sample in a
thermomixer
5. Spin the tube briefly to remove drops from inside
the lid
6. Transfer all the sample to filter column sitting in a
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sample tube
7. Short spin at 500 xg, 1min
8. Take the sample tube for extraction
Chewing gun Use of up to 40 mg of chewing gum cut into small
pieces is recommended
1. Place the chewing-gum sample in a 1.5ml
microcentrifuge tube
2. Add 200μl Buffer BL2 to the sample.
3. Add 20 μl proteinase K, and mix thoroughly by
vortexing for 10 s.
4. Incubate at 56°C for 15 min, vortex-mixing several
times during incubation or place sample in a
thermomixer
5. Spin the tube briefly to remove drops from inside
the lid
6. Take 200μl sample to sample tube for extraction
Cigarette butts Use of approximately 1 cm2 paper from the end of the
cigarette or filter is recommended
1. Place the cigarette-butt sample in a 1.5 ml
microcentrifuge tube
2. Add 200 or 400μl Buffer BL2 to the sample
(Check if the sample has absorbed buffer BL2,if
necessary add more Buffer BL2 to the sample)
3. Add 20 μl proteinase K, and mix thoroughly by
vortexing for 10 s
4. Incubate at 56°C for 15 min, vortex-mixing several
times during incubation or place sample in a
thermomixer
5. Spin the tube briefly to remove drops from inside
the lid
6. Take 200μl sample to sample tube for extraction
Stamps, envelopes Use of a 0.5–2.5 cm2 piece of postage stamp or
envelope is recommended
1. Place all the pieces of sample in a 1.5 ml
microcentrifuge tube
2. Add 200 or 400μ l Buffer BL2 to the sample
(Check if the sample has absorbed buffer BL2,if
necessary add more Buffer BL2 to the sample)
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3. Add 20 μl proteinase K, and mix thoroughly by
vortexing for 10 s
4. Incubate at 56°C for 15 min, vortex-mixing several
times during incubation or place sample in a
thermomixer
5. Spin the tube briefly to remove drops from inside
the lid
6. Take 200μl sample to sample tube for extraction
Controls/ internal control
Using appropriate controls for downstream analysis:
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control (IC) Using a defined quantity control Place in sample tube or the round
well of the reaction chamber
Quality Control In accordance with ZINEXTS’s ISO-certified Quality Management System, each lot of
MagPurix Forensic DNA Extraction Kits is tested against predetermined specifications to
ensure consistent product quality
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MagPurix Viral/Pathogen Nucleic Acids
Extraction Kit A
Cat. No. ZP02011 Process Time: 40-45 minutes (virus)
55-65 minutes (virus+bacteria)
Intended Use
MagPurix Viral/Pathogen Nucleic Acids Extraction Kit A is
used with the MagPurix 12 instrument for extraction of Viral
and bacterial DNA/RNA from cell-free samples, such as
serum, plasma, and other cell-free body fluids
Application
Nucleic acids extracted from MagPurix Viral/Pathogen
Nucleic Acids Extraction kit A can be used in a number of
downstream application including: PCR, qPCR,
Sequencing(NGS), Microarray, RFLP, Southern Blot Analysis
Number Of Tests 48 extractions
Kit Components Kit Contents
(for 48 extractions)
Q’ty
Reagent Cartridge 48 pcs (4x6x2)
Reaction Chamber 48 pcs (4x6x2)
Tip Holder 48 pcs (24x2)
Filtered Tip 50 pcs (25x2)
Piercing Pin 50 pcs (25x2)
Sample Tube (2 mL) 50 pcs (25x2)
Elute Tube (1.5 mL) 50 pcs (25x2)
RNA Carrier (1mg) 1 pcs
Barcode Paper 1 pc
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Reagent Cartridge Content
well-1 Proteinase K solution 40 μl
well-2 Lysis Buffer 1 720 μl
well-3 Binding Buffer 1 1000 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 RNase-free water 1000 μl
well-9 Rnase-free water 1000 μl
well-10 BL2 Buffer 400l
Storage
MagPurix Viral/Pathogen Nucleic Acids Extraction Kit A
should be stored at room temperature (15-25°C). Do
not freeze the reagent cartridges. The Kits are stable for
12 months under the condition
After dissolve the RNA carrier, store it at 4°C
(short-term, up to 1 month) or -20°C (long-term). Do
not freeze–thaw the Frozen RNA carrier more than 3
times. Divide it into conveniently sized aliquots is
recommended
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Starting Material
Sample Type Target Nucleic Acid
Sample Volume (Amount of starting material)
Elution Volume
Serum Total bacterial/
Viral Nucleic Acids
(DNA + RNA)
100-400μl(virus)
100-200(virus/bac
teria)
50-300μl Plasma
CSF
Pretreated Urine
Cell-free body
fluids
Controls/internal
control*
Add controls /internal control in the extraction procedure if the
downstream analysis needed (*see control/internal control on page
76 )
This kit is designed for extraction of viral and bacterial
nucleic acids from plasma or serum, or from a pool of
such cell-free body fluids
After extraction, store the nucleic acid at 4°C (up to
24hours) or -20°C for longer storage. Repeated
freeze–thawing is not allowed
Sample
preparation
The purification procedure is optimized for use with 100-
400 μl serum, plasma*, CSF, pretreated urine or other
cell-free body fluid samples. ( *Blood samples treated
with EDTA or citrate as an anticoagulant can be used for
plasma preparation)
Samples can be either fresh or frozen, provided that they
have not been refrozen after thawing
RNA Carrier (CARRIER) serves two purposes during the
purification procedure. First, it enhances binding of viral
nucleic acids to the silica surface of the magnetic
particles, especially if the sample contains very few
target molecules. Second, the addition of large amounts
of RNA Carrier reduces the chances of RNA degradation
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in the rare event that RNases are not denatured by the
chaotropic salts and detergent in the lysis buffer. If RNA
carrier is not added to the reaction, recovery of DNA or
RNA may be reduced
After collection and centrifugation, plasma, serum, or
CSF can be stored at 2–8°C for up to 6 hours. For longer
storage, we recommend freezing aliquots at –20°C
or –80°C. Thaw samples at room temperature (15–25°C),
and process the samples immediately when they have
equilibrated to room temperature. Do not refreeze the
aliquots after thawing. Repeated freeze–thawing leads to
denaturation and precipitation of proteins, resulting in
reduced viral titers and therefore reduced yields of
nucleic acids. If cryoprecipitates are visible in the
samples, centrifuge at 6800 x g for 3 minutes, transfer
the supernatants to fresh tubes without disturbing the
pellets, and start the purification procedure immediately
RNA Carrier Add 1.0 ml RNase-free water to lyophilized the RNA
Carrier (provided with the kit) and mix by vortexing
Store RNA Carrier at 4°C (short-term, up to 1 month) or
-20°C (long-term). Do not freeze–thaw the frozen carrier
RNA more than 3 times. Divide it into conveniently sized
aliquots is recommended
Before nucleic acid extraction, add RNA Carrier to the
sample is recommended. Add 5 μl Carrier RNA (for 100
μl sample), 10 μl (for 200 μl sample) or 20 μl (for 400 μl
sample) into the Sample Tube
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Controls/ internal control
Using appropriate controls for downstream analysis:
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control(IC) Using a defined quantity control Place in sample tube or the round
well of the reaction chamber
Quality Control
In accordance with ZINEXTS’s ISO-certified Quality Management System, each lot of
MagPurix Viral/Pathogen Nucleic acids Extraction Kit is tested against predetermined
specifications to ensure consistent product quality.
www.ZINEXTS.com Page 78 Vr.5.1
MagPurix Viral/Pathogen Nucleic Acids
Extraction Kit B
Cat. No. ZP02012 Process Time: 55-65 minutes
Intended Use
MagPurix Viral/Pathogen Nucleic Acids Extraction Kit B is used with the MagPurix 12 instrument for extraction of viral and bacterial DNA/RNA from swab samples (cell-rich samples)
Application
Nucleic acids extracted from MagPurix Viral/Pathogen Nucleic Acids Extraction kit B can be used in a number of downstream application including: PCR, qPCR, Sequencing(NGS), Microarray, RFLP, Southern Blot Analysis
Number Of
Tests
48 extractions
Kit components
Kit Contents (for 48 extractions)
Q’ty
Reagent Cartridge 48 pcs (4x6x2)
Reaction Chamber 48 pcs (4x6x2)
Tip Holder 48 pcs (24x2)
Filtered Tip 50 pcs (25x2)
Piercing Pin 50 pcs (25x2)
Sample Tube (2 mL) 50 pcs (25x2)
Elute Tube (1.5 mL) 50 pcs (25x2)
Barcode Paper 1pc
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Reagent Cartridge Content
well-1 Proteinase K solution 40 μl
well-2 Lysis Buffer 2 720 μl
well-3 Binding Buffer 1 720 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 Elution Buffer 1 1000 μl
well-9 Elution Buffer 2 1000 μl
well-10 BL2 Buffer 400L
Storage
MagPurix Viral/Pathogen Nucleic acids Extraction Kit B
should be stored at room temperature (15-25°C). Do not
freeze the reagent cartridges. The Kits are stable for 12
months under the condition
Store the purified Nucleic acids at 4 °C (short- term) or
aliquot and store at -70°C (long-term) before perform the
downstream analysis
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Starting
Material
Bacterial pellet/colony from culture, clinical swab samples
in liquid transport media, environment material (water, soil, etc.) and other cell-rich samples
Use the tissue or paraffin- embedded tissue sections(FFPE) as samples, we recommend to extract DNA by MagPurix Tissue DNA Extraction kit (ZP02004)
The types and amounts of starting material for use in
MagPurix Viral/Pathogen Nucleic Acids Extraction kit B purification procedures are shown in Table listed below,
Sample Type Target Nucleic Acid
Sample volume (Amount of starting material)
Elution Volume
Bacteria Pellet Total Viral/Bacterial Nucleic acids (DNA/RNA)
100-200 μl /Up to 109
bacteria (about OD600 =
3)
50-300 μl
Bacterial colony 100-200 μl /1-3 colony
Swab samples 100-200 μl liquid transport media
Controls/internal control*
Add controls /internal control in the extraction procedure if the downstream analysis needed (* see controls/internal control on page 82)
Sample
Preparation
Sample preparation requirements are highly dependent upon the type of starting material. Due to variations in consistency and viscosity, even similar sample types may require distinct handling
The table below describes the recommendations in processing the primary samples before nucleic acid extraction:
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Table : Preparation of sample material for bacterial nucleic acid extraction
Sample type Procedure
Inactivation the pathogenic microorganism
Recommended pretreatment : Boiling
4. Incubate samples at 95°C for 10 min
5. Centrifuge briefly to collect the complete
sample volume at the bottom of the tube
6. Allow samples to cool down or chill on ice, then
proceeding the following steps according to the
sample type
Viscous samples e.g. BAL, sputum or other mucous specimen
Recommended pretreatment : Liquefaction 1. Prepare a fresh DTT stock solution for
liquefaction * (e.g., 5× conc. DTT stock is about 0.75%)
2. Adjust the final DTT concentration in the sample
to 0.15% by adding DTT stock solution.
3. Incubate the sample (e.g., with shaking at 850r.p.m. for 30 min at 37°C) until it can be pipette easily.
4. Transfer 200 μL to sample tube (Supplied in the kit) * The liquefaction could be done by using other solutions, such as NALC(N-Acetyl-L-Cysteine)
-NaOH or other agents which could digest mucous material
For large volume liquid samples that have low or unknown bacterial loads e.g. urine, water collected from pool/river stream/tower
Recommended pretreatment : Centrifugation 1. Centrifuge the sample for up to 10 min at 20,000
× g to concentrate the bacterial cells in pellet
2. Discard supernatant, resuspend the pellet in 220 μL PBS
3. Take 200 μL to sample tube (Supplied in the kit)
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Swab samples e.g. eye, nasal, pharyngeal, or other swabs
1. Collect samples and place in 1 ml PBS
containing a common fungicide. Incubate for 30min at room temperature
2. Take 200 μL to sample tube
For some gram-positive bacterial species. Especially for samples that contain particles e.g. stool
Recommended pretreatment : Mechanical homogenization
Follow the regular homogenization procedures in
the laboratory.
Bacterial suspension cultures
Take 200 μL culture to sample tube
Bacterial colony 1. Take 1-3 bacterial colony from culture plate with
an inoculation loop and suspend in 220 μL PBS by vigorous stirring
2. Take 200 μL suspension to sample tube
Controls/ internal control
Using appropriate controls for downstream analysis:
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control(IC) Using a defined quantity control Place in sample tube or the round
well of the reaction chamber
Quality Control
In accordance with ZINEXTS’s ISO-certified Quality Management System, each lot of
MagPurix Viral/Pathogen Nucleic acids Extraction Kit is tested against predetermined
specifications to ensure consistent product quality
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MagPurix Viral RNA Extraction Kit
Cat. No. ZP02013 Process Time: 40-45 minutes
Intended Use
MagPurix Viral Nucleic RNA Extraction Kit is used with the
MagPurix 12 instrument for extraction of Viral RNA from
human biological specimens such as serum, plasma, and
other cell-free fluids
Application
Nucleic acids extracted from MagPurix Viral Nucleic Acid
Extraction kit can be used in a number of downstream
application including: PCR, qPCR, Sequencing(NGS),
Microarray, RFLP, Southern Blot Analysis
Number Of Tests
48 extractions
Kit Components
Kit Contents ZP02013-48
Reagent Cartridge 48 pcs (4x6x2)
Reaction Chamber 48 pcs (4x6x2)
Tip Holder 48 pcs (24x2)
Filtered Tip 50 pcs (25x2)
Piercing Pin 50 pcs (25x2)
Sample Tube (2 mL) 50 pcs (25x2)
Elute Tube (1.5 mL) 50 pcs (25x2)
RNA Carrier (1mg) 1pcs
Barcode Paper 1 pc
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Reagent Cartridge Content
well-1 Proteinase K solution 30 μl
well-2 Lysis Buffer 1 720 μl
well-3 Binding Buffer 1 1000 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 RNase-free water 1000 μl
well-9 Rnase-free water 1000 μl
well-10 Empty
Storage
MagPurix Viral RNA Extraction Kit should be stored at
room temperature (15-25°C). Do not freeze the reagent
cartridges. The Kits are stable for 12 months under the
condition
After dissolve the carrier RNA, store it at 4°C
(short-term, up to 1 month) or -20°C (long-term). Do
not freeze–thaw the Frozen carrier RNA more than 3
times
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Starting Material
Sample Type Target Nucleic Acid
Sample Volume (Amount of starting material)
Elution Volume
Serum Total Viral Nucleic
Acids (DNA + RNA)
100-400μl 50-300μl Plasma
CSF
Pretreated Urine
Cell-free body
fluids
Controls/internal
control#
Add controls /internal control in the extraction procedure if the
downstream analysis needed (# see control/internal control on page
86 )
The kit is designed for extraction of viral RNA from
plasma or serum, or from a pool of such cell-free body
fluids
After extraction, store the nucleic acid at 4°C (up to 1
hour) or -20°C for longer storage. Repeated
freeze–thawing is not allowed
Sample
preparation
The purification procedure is optimized for use with 100-
400 μl serum, plasma, CSF, or pretreated urine samples.
( Blood samples treated with EDTA or citrate as an
anticoagulant can be used for plasma preparation)
Samples can be either fresh or frozen, provided that they
have not been refrozen after thawing
After collection and centrifugation, plasma, serum, or
CSF can be stored at 2–8°C for up to 6 hours. For longer
storage, we recommend freezing aliquots at –20°C
or –80°C. Thaw samples at room temperature (15–25°C),
and process the samples immediately when they have
equilibrated to room temperature. Do not refreeze the
aliquots after thawing. Repeated freeze–thawing leads to
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denaturation and precipitation of proteins, resulting in
reduced viral titers and therefore reduced yields of
viralnucleic acids. If cryoprecipitates are visible in the
samples, centrifuge at 6800 x g for 3 minutes, transfer
the supernatants to fresh tubes without disturbing the
pellets, and start the purification procedure immediately
Carrier RNA For RNA virus, adding carrier RNA to the sample before
extraction is recommended !!!
Add 1.0 ml RNase free water to the carrier RNA tube
(provided with the kit) and mix by vortexing. Store it at
4°C (short-term, up to 1 month) or -20°C (long-term). Do
not freeze–thaw the Frozen carrier RNA more than 3
times
Add 5 μl Carrier RNA (for 100 μl sample), 10 μl (for 200 μl
sample) or 20 μl (for 400 μl sample) into the Sample
Tube
Controls/ internal control
Using appropriate controls for downstream analysis:
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control(IC) Using a defined quantity control Place in sample tube or the round
well of the reaction chamber
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Quality Control
In accordance with ZINEXTS’s ISO-certified Quality Management System, each lot of
MagPurix Viral RNA Extraction Kits is tested against predetermined specifications to
ensure consistent product quality.
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MagPurix Plant DNA Extraction Kit
Cat. No. ZP02014 Process Time: 40-45 minutes
Intended Use
MagPurix Plant DNA Extraction Kit is used with the MagPurix 12 instrument for extraction of genomic DNA from plant (leaf, seeds and spores) and fungal tissues. Up to 100 mg of tissue can be used for purification
Application
Nucleic acids extracted from MagPurix Plant DNA Extraction
kit can be used in a number of downstream application
including: PCR, qPCR, Sequencing(NGS), Microarray, RFLP,
Southern Blot Analysis
Number Of Tests
48 extractions
Kit Components
Kit Contents ZP02014-48
Reagent Cartridge 48 pcs (4x6x2)
Reaction Chamber 48 pcs (4x6x2)
Tip Holder 48 pcs (24x2)
Filter Tip 50 pcs
Piercing Pin 50 pcs
Sample Tube (2 mL) 50 pcs
Elute Tube (1.5 mL) 50 pcs
Filter Column 50 pcs
Collection Tube 50 pcs
RNase A(10mg/mL) 1 pc (0.5mL)
Buffer PLA 1 pc (25mL)
Buffer PLB 1 pc (25mL)
Barcode Paper 1 pc
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Reagent Cartridge Content
well-1 Empty
well-2 Lysis Buffer 2 720 μl
well-3 Binding Buffer 1 720 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 Elution Buffer 1 1000 μl
well-9 Elution Buffer 2 1000 μl
well-10 Empty
Storage
MagPurix Plant DNA Extraction Kit should be stored at
room temperature (15-25°C). Do not freeze the reagent
cartridges. The Kits are stable for 24 months under the
condition
Store the purified DNA at 4 °C (short- term) or aliquot and
store at -70°C (long-term) before perform the
downstream analysis
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Starting Material After harvesting plant tissues, it should be frozen in liquid
nitrogen if not be used immediately. It can then be stored
at –80°C.Alternatively; tissue can be dried or lyophilized
after harvesting to allow storage at room temperature
(15–25°C). To ensure DNA quality, samples should be
completely dried within 24 hours of collection
If possible, it is preferable to collect young materials (e.g.,
leaves, needles) since they contain more cells per weight
and therefore result in higher yields
When working with fungi, harvest mycelium directly from
a culture dish or from liquid culture. For liquid culture,
first pellet cells by centrifugation. Remove the
supernatant completely before disruption and lysis.
Fresh, frozen, or freeze-dried fungal material can be use
The disruption method may require optimization to
ensure maximum DNA yield and quality. Complete and
quick disruption of starting material is essential to ensure
high DNA yields and to avoid DNA degradation
Before DNA extraction, plant material is first
mechanically disrupted with lysis buffer (Buffer PLA or
PLB). After Homogenization, remove the debris and other
precipitations by spin through the filter membrane.
Collect the clear flow-through and incubate with RNase A
to digest the RNA in the sample before DNA extraction
Sample preparation Using homogenizer to treat the tissue before extraction is
recommended
For better DNA yield, adding RNase A to sample before
extraction processing
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Sample type Procedure
Plant tissue 1. Add 440l lysis buffer* to sample
2. Perform homogenization by using
proper homogenizer
3. Remove Lysate to filter membrane
sitting in collection tube
4. Short spin at 6000 xg to collect clear
flow-through
5. Add 10l RNase A, incubate for 10min
at room temperature
6. Transfer to sample tube
7. Perform extraction
Yeast A. Suspension culture
1. Centrifuge at 6000xg, 3min
2. Remove supernatant
3. Add 440mL plant lysis buffer*,
vortex mixing 30 sec
4. Transfer 400 L to sample tube
5. Perform extraction
B. Culture colony
3. Take 1-3 colony from culture plate with an inoculation loop and suspend in 440 μL of plant lysis buffer* by vigorous stirring
4. Take 400 μL suspension to sample tube
5. Perform extraction
*We provide two kind plant lysis buffer: PLA and PLB for
dealing with different tissue types. Before extraction a new
tissue type, try these two lysis buffer for getting the optimized
lysis procedure and a better DNA yield
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Controls/ internal control
Using appropriate controls for downstream analysis:
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control(IC) Using a defined quantity control Place in sample tube or the round
well of the reaction chamber
Quality Control In accordance with ZINEXTS’s ISO-certified Quality Management System, each lot of
MagPurix Plant DNA Extraction Kits is tested against predetermined specifications to
ensure consistent product quality
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MagPurix Total RNA Extraction Kit
Cat. No. ZP02015 Process Time: 50-55 minutes
Intended Use
MagPurix Total RNA Extraction Kit is used with the MagPurix
12 instrument for extraction of total RNA from whole blood,
blood cells, animal tissue, plant tissue, yeast or cultured cells
Application
Total RNA extracted from MagPurix Total RNA Extraction
kit can be used in a number of downstream application
including: RT-PCR, qPCR, Sequencing(NGS), Microarray,
Northern blotting
Number Of Tests 36 and 48 extractions
Kit Components Kit Contents Quantity
ZP02015-36 ZP02015-48
Reagent Cartridge 36 pcs (6x6) 48 pcs (4x6x2)
Reaction Chamber 36 pcs (6x6) 48 pcs (4x6x2)
Tip Holder 36 pcs (6X6) 48 pcs (6X4x2)
Filter Tip 38 pcs 50 pcs
Piercing Pin 38 pcs 50 pcs
Sample Tube (2 mL) 76 pcs (38X2) 100 pcs (50X2)
Elute Tube (1.5 mL) 38 pcs 50 pcs
RL A Buffer 25ml 25ml
RL B Buffer 25ml 25ml
Filter column 38 pcs 50 pcs
Collection tube 38 pcs 50 pcs
Barcode Paper 1 pc 1 pc
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Reagent Cartridge Content
well-1 Proteinase K solution 40 μl
well-2 Lysis Buffer 1 720 μl
well-3 Binding Buffer 1 1300 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 RNase-free water 1000 μl
well-9 Rnase-free water 1000 μl
well-10 DNase buffer 1000 μl
Storage
MagPurix Total RNA Extraction Kit should be stored at
room temperature (15-25°C). Do not freeze the reagent
cartridges. The Kits are stable for 15 months under the
condition
After extraction, store RNA at -60 to -80℃
immediately, repeated freeze–thawing is not allowed.
Always handle RNA on ice for downstream analysis
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Purification Protocol
Protocol name Sample vol.
Elute vol.
Description
Total RNA 100-400l
50-200l
Extraction Total RNA and DNA from
sample
Total RNA
(DNA-free)
100-400l
50-200l
Extraction total RNA (DNA-free)* from
sample
* Before use protocol-Total RNA (DNA-free), prepare DNase (not supplied in the kit)
is necessary
Before starting
-Mercaptoethanol (β-ME) must be added to Buffer RLT before use
If performing DNA-free protocol, Prepare DNase before extraction. Place 10 l
DNase in sample tube( at the middle well of sample rack) when extraction
Homogenization is necessary for animal tissue, Plant tissue and yeast before
extraction. Add RL Buffers to sample when perform homogenizing
Wear clean glove, use RNase-free filter tip, and keep work area, pipettors and
reagents free of virus, bacteria and Nuclease contamination
Using RNaseZap® to clean the surface of bench, equipments and pipettors is
one of the most easy way to remove the RNase contaminations of work area
Using RNA stabilized reagent (e.g. RNA later) to treat sample is one of the best
way to protect the RNA if the sample cannot be processing in a RNase-free
working area
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Reagents to be supplied by user
Reagent Description Preparation β -mercaptoethanol
(β -ME)
β -ME reduce disulfide
bonds and irreversibly
denature the RNase and
eliminate RNase released
during cell lysis
Add 10 μl β-ME per 1 ml RL
lysis Buffers*. It can be
stored at RT for up to one
month
Red blood cells lysis
buffer
(RBC lysis buffer)
Lyse Erythrocyte from
whole blood
(Erythrocyte (RBC) lysis
procedure)
10xRBC lysis buffer(100ml)
8.29g NH4Cl (1.5M)
1g KHCO3 (100mM)
0.0372g Na2EDTA (10mM)
Adjust pH7.2-7.4 by HCl
0.2 mm filtered, store for 6
months at 4℃
Dilute 10 times fresh before
use
DNase
To liminate DNA
contamination
Novagen RNase-free DNaseI
(69182-3CN)
*RL lysis buffers means RLA and RLB Buffers. Dispense the β-ME in a fume hood and
wear appropriate protective clothing
Starting Material
Whole Blood
1. Using the fresh whole blood sample for isolation. (within 4 hr, on ice) Freezing
blood is not allowed. The blood sample should be collected in the presence of
an anticoagulant, preferably EDTA, although other anticoagulants such as
citrate, heparin, or ACD (acid citrate dextrose) can also be used.
2. For optimal results, blood samples should be processed within a few hours of
collection and keep in 4℃.
3. Performing Erythrocyte (RBC) lysis procedure before extraction.
4. Using the whole blood samples which have extreme high WBCs no. (more than
10000/l) or concentrated PBMCs( peripheral blood mononucleated cells),
decrease the input volume for extraction is recommended (total WBC no. less
than 5 x 106).
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Tissue
1. To prevent degradation by intracellular RNase, it is important that tissues are
either flash-frozen in liquid nitrogen or stored at –70°C, or processed
immediately following excision.
2. Using RNA stabilized reagent (e.g. RNA later) to treat tissue is another option to
protect the RNA if the sample cannot be froze immediately. Frozen tissue
should not be allowed to thaw during handling (e.g., weighing), keep sample on
ice during cutting or homogenized with RLA Buffer is recommended.
3. After homogenization, using filter column (supplied in the kit) to remove the
insoluble and viscous material of the lysates.
Cells
1. Cells or isolated blood cells can be collected as pellets and either flash-frozen in
liquid nitrogen and stored at –70°C, or processed immediately. Add RLA Buffer
to resuspend pellet for extraction
2. Alternatively, samples can be stored at –70°C in RLA Buffer after disruption and
homogenization. Samples frozen in this way are stable for months.
Plant tissue and yeast
1. Up to 100 mg of sample is first ground in liquid nitrogen or frozen, then add
lysis buffer(RLA or RLB Buffer) to homogenizer
2. Most Plant cells use RLA Buffer for disruption and denaturing sample
3. However, some tissues, such as milky endosperm of maize or mycelia of
filamentous fungi, solidify in RLA Buffer, making the extraction of RNA
impossible. In these cases, RLB Buffer should be used instead.
4. After add lysis buffer(RLA or RLB), samples are place into homogenizer for
homogenization
5. After homogenization, using filter column (supplied in the kit) to remove the
insoluble and viscous material of the lysates.
It is essential to use the correct amount of starting material in order to obtain
optimal RNA yield and purity (as the table below). Use excess quantity is not
helpful in total RNA extraction.
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Sample Type Sample Volume (Amount of starting material)
Elution Volume
Whole blood 200-400 l*
(WBCs no. about 106)
50-200μl**
PBMCs Up to 50 l
(suspended in 200l with RL buffer)
Tissue 10-40mg
(Lysed and suspend with RL buffer)
Cultured cells 200-400 l suspension of primary or
cultured cells (cell no. < 5 x 106)
Plant tissue Up to 100mg
Yeast Up to 100mg
Controls/internal
control
Add controls /internal control in the extraction procedure if the
downstream analysis needed
* Blood cells needs to perform manual RBC lysis procedure
before extraction
** After extraction, store RNA at -60 to -80℃ immediately,
repeated freeze–thawing is not allowed
Sample preparation
Sample Procedure
Whole blood 1. Fresh prepare 1x RBC lysis buffer
2. Add ice-cold two volume RBC lysis buffer to one
volume blood sample
3. Inverting 3-5 times, incubate on ice for 10-15 min
4. Centrifuge at 1000 x g, 10min, 4℃
5. Remove supernatant
6. Resuspend pellet with 220 l RLA Buffer
7. Take 200 l for extraction
PBMCs
(Peripheral Blood
Mononucleated Cells)
1. Resuspend PBMCs with 220l RLA Buffer
2. Vortex mixing for 10 sec
3. Take 200 l for extraction
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Tissue 1. Add 220l RLA Buffer to tissue; make sure the
sample if completely immersed in buffer. Increase
RLA buffer input amount up to 440l if tissue
sample is large.
2. Homogenized tissue by homogenizer
3. Spin down the lysate
4. Remove all the lysate to filter column sitting in
collection tube
5. Centrifuge at 1000 x g , for 5min on 4℃
6. Transfer 200-400l to sample tube
7. Perform extraction
Cultured cells 【Protocol 1】Suspension culture
1. Harvest cell culture
2. Centrifuge at 1000xg, 5min on 4℃
3. Remove supernatant completely
4. Resuspend cell pellet with 22oml RLA Buffer
5. Vortex mixing for 10 sec
6. Take 200l for extraction
【Protocol 2-1】Monolayer culture
1. Trypsinize the cells
2. Harvest the cell in PBS
3. centrifuge at 300xg, 5min on 4℃
4. Remove supernatant
5. Resuspend pellet with 220 l RLA Buffer
6. Vortex mixing for 10sec
7. Take 200l for extraction
【Protocol 2-2】Monolayer culture
1. Scrape the cells with 220-440l RLA Buffer
2. Vortex mixing for 10sec
3. Take 200-400l for extraction
Plant tissue/ Yeast 1. Add 220 -440l RLA or PLB Buffer* to sample, make
sure the sample if complete immersed in buffer
2. Homogenized tissue by homogenizer
3. Remove lysate to filter column sitting in collection
tube
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4. Centrifuge at 1000 x g , for 5min on 4℃
5. Spin down
6. Transfer 200-400l to sample tube
7. Perform extraction
DNA-free RNA
extraction
1. After sample preparation
2. Remove sample to sample tube at top well of
sample rack
3. Place 10 ml DNase in sample tube at the middle well
of sample rack
4. Place elute tube at bottom well of sample rack
5. Select “Total RNA (DNA-free)” protocol to start
extraction
Controls/ internal control
Using appropriate controls for downstream analysis:
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control(IC) Using a defined quantity control Place in the round well of the reaction
chamber
Quality Control
In accordance with ZINEXTS’s ISO-certified Quality Management System, each lot of
MagPurix Total RNA Extraction Kit is tested against predetermined specifications to
ensure consistent product quality.
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MagPurix Viral Nucleic Acid Extraction Kit 800
Cat. No. ZP02016 Process Time: 55-65 minutes
Intended Use
MagPurix Viral Nucleic Acid Extraction Kit 800 is used with
the MagPurix 12 instrument for extraction of Viral DNA or
RNA from human biological specimens such as serum,
plasma, and other cell-free fluids.
Application
Nucleic acids extracted from MagPurix Viral Nucleic Acid
Extraction kit can be used in a number of downstream
applications including: PCR, qPCR, Sequencing(NGS),
Microarray, RFLP, Southern Blot Analysis
Number Of Tests
48 extractions
Kit Components
Kit Contents ZP02016-48
Reagent Cartridge 48 pcs (4x6x2)
Reaction Chamber 48 pcs (4x6x2)
Tip Holder 48 pcs (24x2)
Filter Tip 50 pcs
Piercing Pin 50 pcs
Sample Tube (2 mL) 50 pcs
Elute Tube (1.5 mL) 50 pcs
RNA Carrier (1mg) 1 pcs
Barcode Paper 1 pc
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Reagent Cartridge Content
well-1 Proteinase K solution 40 μl
well-2 Lysis Buffer 1 1000 μl
well-3 Binding Buffer 1 1600 μl
well-4 Magnetic Bead Solution 800 μl
well-5 Washing Buffer 1 1000 μl
well-6 Washing Buffer 2 1000 μl
well-7 Washing Buffer 3 1000 μl
well-8 RNase-free water 1000 μl
well-9 Rnase-free water 1000 μl
well-10 Empty
Storage
MagPurix Viral Nucleic Acid Extraction Kit 800 should be
stored at room temperature (15-25°C). Do not freeze
the reagent cartridges. The Kits are stable for 12
months under the condition
After dissolve the carrier RNA, store it at 4°C
(short-term, up to 1 month) or -20°C (long-term). Do
not freeze–thaw the Frozen carrier RNA more than 3
times
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Starting Material
Sample Type Target Nucleic Acid
Sample Volume (Amount of starting material)
Elution Volume
Serum Total Viral Nucleic
Acids (DNA + RNA)
600-800μl 50-300μl Plasma
CSF
Pretreated Urine
Cell-free body
fluids
Controls/internal
control#
Add controls /internal control in the extraction procedure if the
downstream analysis needed (# see control/internal control on page
24 )
The kit is designed for extraction of viral nucleic acids
(e.g., those of HIV, HCV, HBV, or HPV) from plasma or
serum, or from a pool of such cell-free body fluids
After extraction, store the nucleic acid at 4°C (up to
24hours) or -20°C for longer storage. Repeated
freeze–thawing is not allowed
Sample
preparation
The purification procedure is optimized for use with 100-
400 μl serum, plasma, CSF, or pretreated urine samples.
( Blood samples treated with EDTA or citrate as an
anticoagulant can be used for plasma preparation)
Samples can be either fresh or frozen, provided that they
have not been refrozen after thawing
After collection and centrifugation, plasma, serum, or
CSF can be stored at 2–8°C for up to 6 hours. For longer
storage, we recommend freezing aliquots at –20°C
or –80°C. Thaw samples at room temperature (15–25°C),
and process the samples immediately when they have
equilibrated to room temperature. Do not refreeze the
aliquots after thawing. Repeated freeze–thawing leads to
denaturation and precipitation of proteins, resulting in
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reduced viral titers and therefore reduced yields of
viralnucleic acids. If cryoprecipitates are visible in the
samples, centrifuge at 6800 x g for 3 minutes, transfer
the supernatants to fresh tubes without disturbing the
pellets, and start the purification procedure immediately
Carrier RNA For RNA virus, adding carrier RNA to the sample before
extraction is recommended !!!
Add 1.0 ml RNase free water to the carrier RNA tube
(provided with the kit) and mix by vortexing. Store it at
4°C (short-term, up to 1 month) or -20°C (long-term). Do
not freeze–thaw the Frozen carrier RNA more than 3
times
Add 5 μl Carrier RNA (for 100 μl sample), 10 μl (for 200 μl
sample) or 20 μl (for 400 μl sample) into the Sample
Tube
Controls/ internal control
Using appropriate controls for downstream analysis: :
Type Description Location
Positive control Using sample which positive for
target
Place in sample tube
Negative control Using sample which negative for
target or water(NTC)
Place in sample tube
Internal control(IC) Using a defined quantity control Place in sample tube or the round well of
the reaction chamber*
*Use RNA as internal control, add it into round well of reaction chamber is recommended
Quality Control In accordance with ZINEXTS’s ISO-certified Quality Management System, each lot of
MagPurix Viral Nucleic Acid Extraction Kits 800 is tested against predetermined
specifications to ensure consistent product quality
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Protocol of extraction
1. Turn the power switch on and waiting for the LCM
screen turn on and shows “MagPurix 12 System
Stand-By”.
2. Press “Start” button
(The system will process self-testing, and then go to
steady mode)
Note:
The system will block main functions before the
completion of self-testing process.
3. Open the sliding door and remove the sample rack
from the instrument.
4. Load Reagent Cartridges, and all plastics disposables
(Reaction Chamber, Tip Holder, Piercing Pin, Filtered
Tip and Pestle (optionally supplied with some kit types)
Insert the cartridges
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How to pull apart reagent cartridges
Slash open the dotted line with nail and snap it with a little bit
force.
Insert Reaction Chambers
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Insert Tip Holder
Insert Piercing Pins
Insert Filtered Tips
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Note:
1. The positions of piercing pin and filtered tip; the 2nd
one should be “EMPTY”.
2. Load one Reagent Cartridge and one set of plastic
disposable per sample.
Important:
- Set Cartridges in the order of the number from
left to right.
- Make sure that Cartridges are inserted in to the
Cartridge Tray tightly.
- You can load 1-12 cartridges on the tray
depending on the number of samples that you
wish to process.
5. Load Sample Tube and Elute Tube to Sample Rack on the
bench
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Insert Sample Tube on to Sample Rack
Insert Elute Tube on to Sample Rack
6. Load the sample(s) to Sample Tube
Note:
- Pretreatments are essential for some sample
types before loading to Sample Tube. Please refer
to the handbook of reagent kits for details.
- Make sure the caps of Elute Tube are open as the
figure shown above.
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7. Place Sample Rack on the instrument platform
Note:
- Use two hands to handle the Sample Tray.
- Make sure the Sample Tray be placed correctly on to
the instrument
8. Close the door
9. Scan the protocol barcodes to select purification protocol,
sample volume and elute volume
Note:
- There is one protocol barcode paper enclosed in
the reagent kit box
- Protocol’s name, sample volume and elution
volume will be shown on LCM screen after
protocol barcode is scanned
10. Follow the instructions displayed on LCM screen to double
check the operating steps being completed before
program running.
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11. Push “Enter” to confirm. Instrument will start to run the
protocol program automatically until whole processes are
completed.
Note:
- It takes from 30 to 45 minutes to complete the
extraction according to reagent types
12. At the end of the run, the instrument beeps briefly and the
LCM shows “Protocol Completed”
13. Open the instrument door
14. Remove the elute tubes containing the purified nucleic
acid
Note: Store the purified nucleic acids at 4℃ for
short-term storage or store at -70℃ for long-term
storage
15. Discard the used cartridges, all plastic consumables into
biohazard waste. Do not reuse the cartridges
16. If you’re not using the instrument, place the Sample Rack
back to workplace, close the instrument door and push
“Start” button for 2 secs to get into “sleeping mode”.
And for longer time not using the instrument turn the
power switch off.
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Zinexts Life Science Corporation
www.zinexts.com
2F.-2, No.122, Qiaohe Rd., Zhonghe Dist.,
New Taipei City 235, Taiwan
Tel: +886 2 2246 3579
Fax: +886 2 2243 8570
Mail: [email protected]