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USB® ExoSAP-IT® PCR Product Cleanup
For standard and high-throughput platforms - now available in a faster formulation
ExoSAP-IT®
PCR Cleanup reagent.The gold standard.
USB® ExoSAP-IT® PCR Product Cleanup means the ease of one-step purification.
The ExoSAP-IT method is a unique, one-tube, one-step, enzymatic cleanup reagent for PCR product cleanup.
100% sample recovery – no loss of PCR products regardless of the fragment size
Removes excess primers and dNTPs – does not interfere with downstream applications
Superior results with ExoSAP-IT reagent – improve accuracy with higher yields and
full PCR product recovery
Cited in over 7,000 publications – the enzymatic PCR cleanup solution used by leading scientists
Cuts protocol time in half - faster cleanup using automated platforms and multi-channel pipettes
USB ExoSAP-IT PCR Product Cleanup – available for standard and high-throughput, automated platforms
One step to superior sequencing results.
One-step enzymatic cleanup of PCR products
The ExoSAP-IT method is a unique, one-step enzymatic cleanup of PCR
products that eliminates unincorporated primers and dNTPs so they
do not interfere with downstream sequencing or SNP analysis. The
ExoSAP-IT cleanup reagent, based on Exonuclease I and Shrimp Alkaline
Phosphatase (SAP) is added directly to the PCR product to degrade
primers and dephosphorylate dNTPs that were not consumed in the
reaction (Fig. 1). Since ExoSAP-IT reagent is active in commonly used
PCR buffers, no buffer exchange is required. Treatment is carried out for
15 minutes at 37°C and is followed by a 15 minute incubation period
at 80°C to completely inactivate both enzymes. PCR products are then
immediately ready for downstream sequencing reactions without any
additional manipulation.
One-tube/one-step PCR cleanup – Add ExoSAP-IT reagent directly
to PCR product
Eliminate spin columns – Decrease time and expense while
increasing yield
Conserve PCR samples – 100% recovery of both short and long
PCR products for accurate sequencing results (Fig. 2)
Fig. 2. Achieve high data quality from PCR products
Fig. 1. ExoSAP-IT PCR Product Treatment Overview
PCR Mixture Post-ampli�cation
PCR Product
+
Nucleosides
+
Inorganic Phosphate (Pi)
Excess Primers Excess dNTPs
37°C, 15 min for treatment
80°C, 15 min to inactivate
Add ExoSAP-IT
reagent
Stable at 25°C for 8 hours – Retains full functional activity
and is stable at 4°C for one week
Green option – Generate less waste with our single-tube solution to
PCR purification vs. columns
Fluorescent sequencing results of a 100 bp
pUC18 PCR fragment sequenced with a -20
Fwd primer using fluorescent sequencing
reagents. PCR clean-up performed with:
(top) ExoSAP-IT reagent; (bottom) a column
designed for PCR cleanup. Base miscalls in
(bottom) are due to inherently low yields of
short PCR products when using columns.
ExoSAP-IT PCR Product Cleanup
78250
78200
78201
78202
78205
20 reactions
100 reactions
1500 reactions
2,000 reactions
5,000 reactions
USB HT ExoSAP-IT High-Throughput reagents are an alternative
formulation of ExoSAP-IT PCR Product Cleanup specifically designed
for the unique requirements of high-throughput, automated platforms
and multi-channel pipettes. New to our family of patented ExoSAP-IT
products, HT ExoSAP-IT Fast High-Throughput PCR Cleanup reagent
quickly provides accurate and consistent results in high-throughput
applications. HT ExoSAP-IT Fast reagent is over 50% faster than
standard HT ExoSAP-IT reagent with a 14 minute total protocol time.
HT ExoSAP-IT reagent has a decreased viscosity for robotic pipetting,
yet maintains the same convenience and stability that you have come
to expect from the ExoSAP-IT method.
HT ExoSAP-IT Fast reagent
50% faster cleanup protocol
Stable at 4°C for one month
Scalable for PCR cleanup from a single tube up to a 384-well
plate PCR reaction
High quality—Accurate results
In comparison to competitor products, HT ExoSAP-IT reagent allows
for longer read lengths and greater confidence in the accuracy of those
reads than did the alternative, AMPure® XP beads.
PCR products cleaned up with HT ExoSAP-IT Fast reagent prior to
automated sequencing displayed superior sequencing results compared
to the untreated equivalent (Fig. 3).
Now available in a faster high-throughput formulation
HT ExoSAP-IT High-Throughput PCR Product Cleanup
78395 480 reactions x 8-tube strip
5,760 reactions x 1 plate (12 x 8-tube strips)
23,040 reactions - 4 plates
1,000 reactions (2 ml)
5,000 reactions (10 ml)
HT ExoSAP-IT Fast High-Throughput PCR Product Cleanup
78595 20 reactions
480 reactions x 8-tube strip
5,760 reactions x 1 plate (12 x 8-tube strips)
23,040 reactions x 4 plates
1,000 reactions - 2 ml
5,000 reactions - 10 ml
Fig. 3. High quality sequencing data after treatment with HT ExoSAP-IT Fast reagent
HTExoSAP-ITFast
reagentTreated
Untreated
Sequencing of a 1 kb PCR product cleaned with HT ExoSAP-IT Fast reagent (top) or untreated (bottom). HT ExoSAP-IT Fast treatment prior to sequencing
eliminates miscalls and improves sequencing quality scores (numbers and bars above sequence; >50, probability of error ≤0.001). Sequence shown is
within the first 150 bases.
For research use only. Not for use in diagnostic procedures.Affymetrix, USB, ExoSAP-IT, and VeriQuest are registered trademarks of Affymetrix, Inc. All other trademarks are the property of their respective owners. ExoSAP-IT for PCR Product Cleanup, HT ExoSAP-IT High-Throughput PCR Product Cleanup, and HT ExoSAP-IT Fast High-Throughput PCR Product Cleanup—These products are covered by patents owned by Affymetrix, including US Patent Nos. 6,379,940 and 6,387,634 and corresponding patents in Canada and Japan. ExoSAP-IT for PCR Product Cleanup, HT ExoSAP-IT High-Throughput PCR Product Cleanup and HT ExoSAP-IT Fast High- Throughput PCR Product Cleanup—These products or portions thereof are sold under license from GE Healthcare under US Patent Nos. 5,741,676 and 5,756,285. HotStart-IT Taq DNA Polymerase – Methods for using this product may be covered by US Patent No. 7,700,281. SYBR is a registered trademark of Molecular Probes, Inc. and is provided under an agreement with Molecular Probes, Inc. Agencourt and AMPure are registered trademarks of Agencourt.
End Point PCR
Standard
71160 Taq DNA Polymerase
71162 Taq PCR Master Mix (2X)
71190 RubyTaqTM DNA Polymerase
71191 RubyTaq PCR Master Mix (2X)
Long and accurate PCR
71180 FideliTaqTM DNA Polymerase
71182 FideliTaq PCR Master Mix (2X)
71155 HotStart-IT® FideliTaq DNA Polymerase
71156 HotStart-IT FideliTaq PCR Master
Mix (2X)
Hot start
71195 HotStart-IT Taq DNA Polymerase
71196 HotStart-IT Taq PCR Master Mix (2X)
71194 HotStart-IT Binding Protein
Hot start multiplex
MP1100 Promoter Methylation PCR Kit
71199 MagniTaqTM Multiplex PCR
Master Mix (2X)
Real-Time qPCR
Components & kits
75780 First-Strand cDNA Synthesis Kit for
Real-Time PCR
71170 VeriQuest® Taq DNA Polymerase
SYBR
75600 VeriQuest SYBR® Green qPCR Master
Mix (2X)
75665 VeriQuest SYBR Green qPCR Master
Mix with Fluorescein (2X)
75690 VeriQuest Fast SYBR Green qPCR
Master Mix (2X)
75675 VeriQuest Fast SYBR Green qPCR
Master Mix with Fluorescein (2X)
75762 HotStart-IT SYBR Green qPCR
Master Mix (2X)
75760 HotStart-IT SYBR Green qPCR
Master Mix with UDG (2X)
Probe
75650 VeriQuest Probe qPCR Master Mix
(2X)
75660 VeriQuest Probe qPCR Master Mix,
No Reference Dye (2X)
75680 VeriQuest Fast Probe qPCR Master
Mix (2X)
75685 VeriQuest Fast Probe qPCR Master
Mix, No Reference Dye (2X)
75766 HotStart-IT Probe qPCR Master Mix
(2X)
75764 HotStart-IT Probe qPCR Master Mix
with UDG (2X)
Real-Time Reverse Transcription
PCR (qRT-PCR)
One-step master mixes
75705 VeriQuest SYBR Green One-Step
qRT-PCR Master Mix (2X)
75715 VeriQuest SYBR Green
One-Step qRT-PCR Master Mix with
Fluorescein (2X)
75700 VeriQuest Probe One-Step
qRT-PCR Master Mix (2X)
75710 VeriQuest Probe One-Step
qRT-PCR Master Mix, No Reference
Dye (2X)
75770 HotStart-IT SYBR Green One-Step
qRT-PCR Master Mix Kit
75772 HotStart-IT Probe One-Step
qRT-PCR Master Mix Kit
Kits
75780 First-Strand cDNA Synthesis Kit
for Real-Time PCR
Reverse Transcription RT-PCR
One-step master mix
78370 RT-PCR Master Mix (2X)
71185 FideliTaq RT-PCR Master Mix (2X)
Kits
78350 One-Step RT-PCR Kit
78355 Two-Step RT-PCR Kit
PCR Components
Nucleotides
77212 10 mM PCR Nucleotide Mix
77119 25 mM PCR Nucleotide Mix
77330 10 mM PCR Nucleotide Mix
with dUTP
Reagents
77507 Betaine, 5 M Solution
71786 Nuclease-Free Water
Affymetrix, Inc.
USB® Products
26111 Miles Road
Cleveland, Ohio 44128
Tel: 800-321-9322 | 216-765-5000
Fax: 800-535-0898 | 216-464-5075
usb.affymetrix.com
Affymetrix UK Ltd.
USB® Products
Voyager, Mercury Park,
Wycombe Lane, Wooburn Green,
High Wycombe HP10 0HH, UK
Tel: +44 (0)1628 55 2600
Fax: +44 (0)1628 55 2675
Affymetrix Pte Ltd
USB® Products
7 Gul Circle, #2M-01
Keppel Logistics Building
Singapore 629563
Tel: +65 63957301
Fax: +65 63957300
© 2015 Affymetrix, Inc. All rights reserved. Rev 03/23/15P/N USB05033 Rev. 1
Related products
For research use only. Not for use in diagnostic procedures.
Affymetrix, USB and ExoSAP-IT are registered trademarks of Affymetrix, Inc.
ExoSAP-IT PCR Product Cleanup is covered by patents owned by Affymetrix, including US Patent Nos. 6,379,940 and 6,387,634 and corresponding patents in Canada and Japan.
ExoSAP-IT PCR Product Cleanup—This product or portions thereof are sold under license from GE Healthcare under US Patent Nos. 5,741,676 and 5,756,285.
© 2014 Affymetrix, Inc. All rights reserved.
Affymetrix, Inc. usb.affymetrix.comUSA: [email protected] P 78200BEurope: [email protected] rev 06/14
USB® ExoSAP-IT® PCR Product CleanupProduct numbers 78200/01/02/05/50Brief protocol
ExoSAP-IT reagent treats PCR products ranging in size from less than 100 bp to over 20 kb with absolutely no sample loss by removing unused primers and nucleotides.
Add ExoSAP-IT reagent directly to the reaction products following PCR. ExoSAP-IT PCR Product Cleanup is active in commonly used PCR buffers, so no buffer exchange is required. After treatment, ExoSAP-IT reagent is inactivated by heating to 80°C for 15 minutes. The treated PCR products are now ready for subsequent analysis in applications that require DNA to be free of excess primers and nucleotides.
PCR cleanup protocol:1. Remove ExoSAP-IT reagent from -20°C freezer and keep
on ice throughout this procedure.2. Mix 5 μl of a post-PCR reaction product with 2 μl of
ExoSAP-IT reagent for a combined 7 μl reaction volume. Note: When treating PCR product volumes greater than
5 μl, simply increase the amount of ExoSAP-IT reagent proportionally.
3. Incubate at 37°C for 15 minutes to degrade remaining primers and nucleotides.
4. Incubate at 80°C for 15 minutes to inactivate ExoSAP-IT reagent.
5. The PCR product is now ready for use in DNA sequenc-ing, SNP analyses, or other primer-extension applications. Treated PCR products may be stored at -20°C until required.
Note: Store ExoSAP-IT reagent in a non-frost-free freezer.
USB® HT ExoSAP-IT® High-Throughput PCR Cleanup
USB ExoSAP-IT cleanup reagent, which revolutionized PCR
cleanup for general sequencing, extends the revolution to high-
throughput sequencing with HT ExoSAP-IT reagent. HT ExoSAP-IT
reagent is an alternative formulation of ExoSAP-IT reagent specif-
ically designed for the unique requirements of high-throughput,
automated platforms. HT ExoSAP-IT reagent has a decreased vis-
cosity for robotic pipetting yet maintains the same convenience
and stability that you have come to expect from the ExoSAP-IT
products. Like ExoSAP-IT, HT ExoSAP-IT reagent is a mixture of
Exonuclease I and Shrimp Alkaline Phosphatase, Recombinant for
the enzymatic removal of excess primers and dNTPs following a
PCR reaction. Also, like ExoSAP-IT reagent, HT ExoSAP-IT reagent
allows PCR cleanup with a single pipetting step. HT ExoSAP-IT
reagent is scalable for PCR cleanup in a single tube up to a 384-
well plate PCR reaction.
Features and benefits
One-tube PCR cleanup—Add HT ExoSAP-IT reagent directly
to PCR product
100% sample recovery—No loss of PCR products regard-
less of the fragment size
Simple, single-step—Replaces the multiple steps and
wasted time inherent with beads or columns (see table)
High-throughput processing—Low viscosity formulation
allows robotic pipetting
Removes excess primers and dNTPs—Does not interfere
with downstream applications
• sequencing
• SNP analysis
• single base extension
• fragment analysis
Scalable—Treat reaction volumes from 5 µl to 5 L
Convenient packaging—Available in 8-tube strips and
96-well formats
Stable at 25°C for 8 hours—Retains full functional activity
and is stable for one week at 4°C
High quality – accurate results
HT ExoSAP-IT High-Throughput PCR Cleanup reagent is designed
to provide accurate and consistent results in high-throughput
applications. Identical PCR products were purified with
HT ExoSAP-IT reagent and with AMPure® XP beads, and the
treated PCR products were then sequenced. The results
highlight the robust and accurate data obtainable with
HT ExoSAP-IT reagent. A 1,007 bp PCR fragment from human
jagged1 was amplified from 15 ng of human genomic DNA in a
100 μl reaction using HotStart-IT® FideliTaq™ PCR Master Mix
(PN 71156). A sample of 5 μl of the PCR reaction was treated
with HT ExoSAP-IT reagent and 40 μl of PCR reaction was
processed with Agencourt® AMPure XP beads, both according to
the manufacturer’s directions.
Read lengths from PCR products treated with HT ExoSAP-IT
reagent were on average 51 bases longer than those from PCR
products purified with AMPure XP beads. In addition, the
HT ExoSAP-IT-treated PCR products had higher Phred 20 scores
than the AMPure XP purified PCR products (Table 1). Thus,
HT ExoSAP-IT reagent allowed longer read lengths and greater
confidence in the accuracy of those reads than did AMPure XP
beads. Sequencing also revealed that the bead-purified DNA
had base miscalls, which is likely an effect of sample loss.
HT ExoSAP-IT-treated samples showed no miscalls and had
100% recovery (Fig. 1).
Table 1. HT ExoSAP-IT reagent gives better sequencing results.
HT ExoSAP-IT reagent
AMPure XP beads
Number bases read (Average of 16 samples)
901 850
Total miscalls 0 25
Pass rate (%)* 100% 99.7%
Phred 20** 822 ± 9 776 ± 83
* Pass rate – Average Phred value greater than 20 bases between 100 bp and 300 bp.
** Phred 20 values over the entire length of sequence (no trimming ~970 bp)
Fig. 1. Seqencing of a 1,007 bp treated PCR product
A 1,007 bp fragment was amplified and treated with HT ExoSAP-IT
reagent (above) or Agencourt AMPure XP beads (below) and sequenced.
Chromatograms revealed no miscalls with HT ExoSAP-IT reagent but three
miscalls with Agencourt AMPure XP beads at position 203, 204 and 220.
HT ExoSAP-IT
AMPure XP
3 miscalls
Affymetrix, Inc.
USB® Products
26111 Miles Road
Cleveland, Ohio 44128
Tel: 800-321-9322 | 216-765-5000
Fax: 800-535-0898 | 216-464-5075
usb.affymetrix.com
Affymetrix UK Ltd.
USB® Products
Voyager, Mercury Park,
Wycombe Lane, Wooburn Green,
High Wycombe HP10 0HH, UK
Tel: +44 (0)1628 55 2600
Fax: +44 (0)1628 55 2675
Affymetrix Pte Ltd
USB® Products
7 Gul Circle, #2M-01
Keppel Logistics Building
Singapore 629563
Tel: +65 63957301
Fax: +65 63957300
For research use only. Not for use in diagnostic procedures. Affymetrix, USB, ExoSAP-IT and HotStart-IT are registered trademarks of Affymetrix, Inc. FideliTaq is a trademark of Affymetrix, Inc. Exonuclease I/Shrimp Alkaline Phosphatase—This product is licensed under US Patent Nos. 5,741,676 and 5,756,285, and corresponding patents issued in other countries. Purchase of this product includes a license to use this product in a restricted way within the scope of rights granted to USB by GE Healthcare. No other license is granted to the purchaser either directly or by implication, estoppel or otherwise. ExoSAP-IT is covered by US Patent Nos. 6,379,940 and 6,387,634. Agencourt and AMPure are registered trademarks of Agencourt. All trademarks are the property of their respective owners.
100% recovery from a range of fragment sizes with
HT ExoSAP-IT reagent
HT ExoSAP-IT reagent ensures 100% recovery and provides
effective cleanup of all amplicon sizes. In contrast, AMPure XP
beads were ineffective at purifying small amplicons whether
determined by image analysis (Fig. 2) or by PicoGreen assay
(Table 2).
Table 2. HT ExoSAP-IT reagent gives 100% recovery.
PCR size
DNA recovery after purification with
Agencourt AMPure XP beads
DNA recovery after purification with
HT-ExoSAP-IT reagent
86 bp 10% 100%
103 bp 12% 100%
545 bp 63% 100%
1,007 bp 88% 100%
HT ExoSAP-IT-treated PCR product is stable
HT ExoSAP-IT reagent is rigorously tested and subject to strict
quality control. This helps to insure the integrity of the treated
PCR product. Fig. 3 demonstrates that no product degradation
occurred after storage of HT ExoSAP-IT-treated PCR product for
one week at 25°C.
HT ExoSAP-IT reagent
Product code Pack size
78395 1 each – 480 reactions x 8-tube strip
1 pack – 5,760 reactions x 1 plate (12 x 8-tube strips)
4 pack – 23,040 reactions x 4 plates
1,000 reactions – 2 ml
5,000 reactions – 10 ml
Fig. 3. High stability of HT ExoSAP-IT-treated products
A 1,007 bp DNA fragment amplified from genomic DNA with HotStart
FideliTaq PCR Master Mix (PN 71156) was treated with HT ExoSAP-IT
reagent and stored for seven days at: -20ºC (lane 1), 4ºC (lane 2), or 25ºC
(lane 3). Samples were visualized on 1.5% agarose/TAE/ethidium bromide
gel. Lane MW: DNA Ladder, 1 kb Plus (PN 76714).
1 kb -20°C 4°C 25°C
Fig. 2. Greater recovery of PCR product with HT ExoSAP-IT reagent
Equivalent volumes of PCR product were visualized on an ethidium bromide agarose gel and the band volume determined using image analysis. HT-ExoSAP-IT
reagent allowed recovery of 100% of the 86 and 103 bp PCR products but AMPure XP beads allowed only 8 and 14% recovery, respectively. Un – Untreated,
HT – HT-ExoSAP-IT treated, XP – AMPure XP purified, 100 bp – Affymetrix 100 bp ladder, bands from 100 to 1,000 bp by 100 bp intervals (PN 76712).
© 2015 Affymetrix, Inc. All rights reserved. Rev 01/01/15P/N USB05020 Rev. 1
HT ExoSAP-IT® Fast High-Throughput PCR Product Cleanup
From the leader in enzymatic PCR cleanup comes a
new, faster option. Introducing HT ExoSAP-IT Fast for High-
Throughput PCR Product Cleanup: a faster version of our flagship
ExoSAP-IT reagent.
Like the standard formulation, HT ExoSAP-IT Fast reagent is
a mixture of Exonuclease I and Shrimp Alkaline Phosphatase for
the enzymatic removal of excess primers and dNTPs following a
PCR reaction. When PCR amplification is complete, any uncon-
sumed dNTPs and primers remaining in the PCR product mixture
will interfere with downstream applications. The ExoSAP-IT
products remove these contaminants. HT ExoSAP-IT Fast reagent
is scalable for PCR cleanup from a single tube up to a 384-well
plate PCR reaction.
Why HT ExoSAP-IT Fast reagent?
Half the time of standard enzymatic protocols
100% recovery and only 5 μl of PCR product needed
One simple pipetting step
Formulated and formatted for automated platforms and
multi-channel pipettes
The most stable ExoSAP-IT reagent available
Adding to our family of patented ExoSAP-IT products,
HT ExoSAP-IT Fast reagent quickly provides accurate and consis-
tent results in high-throughput applications. HT ExoSAP-IT Fast
reagent is over 50% faster than standard HT ExoSAP-IT reagent
with a 14 minute total protocol time. Add it directly to the PCR
product and incubate at 37°C for just 7 minutes. After PCR
treatment, inactivate it by simply heating to 80°C for another 7
minutes (Fig. 1).
This product is ideal for high volume labs that require quick
sample turn around. With such a brief and simple protocol,
proper sample cleanup never needs to be sacrificed due to pro-
cessing or instrument time constraints. Ensure the highest quality
Sanger sequencing results while keeping sample processing mov-
ing quickly and efficiently (Fig. 2).
HT ExoSAP-IT Fast reagent has a decreased viscosity yet main-
tains the same convenience and stability that you have come to
expect from the ExoSAP-IT products. As with all products in the
ExoSAP-IT product line, HT ExoSAP-IT Fast reagent allows PCR
cleanup with a single pipetting step. HT ExoSAP-IT Fast reagent
is scalable for PCR cleanup from a single tube up to a 384-well
plate PCR reaction. PCR products cleaned up with HT ExoSAP-IT
Fast reagent prior to automated sequencing displayed superior
sequencing results compared to the untreated equivalent (Fig. 3).
HT ExoSAP-IT Fast reagent displayed excellent stability in all
formats tested including vials, 8-tube strips, and 96-well plates.
With excellent stability and a greater than 50% faster protocol,
HT ExoSAP-IT Fast reagent is ideal for high-throughput assays
where robotic platforms are utilized and large volumes of sam-
ples are processed in successive testing runs.
Collectively, these results clearly demonstrate the effective-
ness of HT ExoSAP-IT Fast reagent to ensure the highest quality
sequences while keeping sample processing moving quickly
and efficiently.
Fig. 1. HT ExoSAP-IT Fast High-Throughput PCR Product Cleanup overview
Affymetrix, Inc.
USB® Products
26111 Miles Road
Cleveland, Ohio 44128
Tel: 800-321-9322 | 216-765-5000
Fax: 800-535-0898 | 216-464-5075
usb.affymetrix.com
Affymetrix UK Ltd.
USB® Products
Voyager, Mercury Park,
Wycombe Lane, Wooburn Green,
High Wycombe HP10 0HH, UK
Tel: +44 (0)1628 55 2600
Fax: +44 (0)1628 55 2675
Affymetrix Pte Ltd.
USB® Products
7 Gul Circle, #2M-01
Keppel Logistics Building
Singapore 629563
Tel: +65 63957310
Fax: +65 63957300
For Research Use Only. Not for use in diagnostic procedures. Affymetrix, USB, and ExoSAP-IT are registered trademarks of Affymetrix, Inc. All other trademarks are the property of their respective owners. ExoSAP-IT for PCR Product Cleanup, HT ExoSAP-IT High-Throughput PCR Product Cleanup, and HT ExoSAP-IT Fast High-Throughput PCR Product Cleanup—These products are covered by patents owned by Affymetrix, including US Patent Nos. 6,379,940 and 6,387,634 and corresponding patents in Canada and Japan. ExoSAP-IT for PCR Product Cleanup, HT ExoSAP-IT High-Throughput PCR Product Cleanup and HT ExoSAP-IT Fast High-Throughput PCR Product Cleanup—These products or portions thereof are sold under license from GE Healthcare under US Patent Nos. 5,741,676 and 5,756,285.
© 2015 Affymetrix, Inc. All rights reserved. Rev 09/30/15P/N USB05032 Rev. 2
HT ExoSAP-IT Fast High-Throughput PCR Product Cleanup
Product code Pack size
78595 20 reactions
1 each – 480 reactions x 8-tube strip
1 pack – 5,760 reactions x 1 plate (12 x 8-tube strips)
4 pack – 23,040 reactions x 4 plates
1,000 reactions – 2 ml
5,000 reactions – 10 ml
Fig. 3. High quality sequencing data after treatment with HT ExoSAP-IT Fast reagent
Sequencing of a 1 kb PCR product cleaned with HT ExoSAP-IT Fast reagent (top) or untreated (bottom). HT ExoSAP-IT Fast treatment prior to sequencing elimi-
nates miscalls and improves sequencing quality scores (numbers and bars above sequence; >50, probability of error ≤0.001). Sequence shown is within the first
150 bases.
HTExoSAP-ITFast
reagentTreated
Untreated
Fig. 2. Hands-on vs. hands-off time savings
USB® ExoSAP-IT® PCR Product Cleanup
The gold standard in enzymatic PCR cleanup
Fast track your sequencing results
ExoSAP-IT®
PCR Cleanup ReagentsOne step to superior sequencing results
Our one-step ExoSAP-IT Express reagent ensures quality
sequencing results in a fraction of the time.
5 minute protocol—Fastest turnaround time to results
One-tube, one-step PCR cleanup—Add directly to PCR
product
Novel enzyme technology—Enzymes are irreversibly
inactivated in just 1 minute at 80°C
Conserve PCR samples—100% recovery and no loss of PCR
products, regardless of amplicon length
Scaleable—Treat PCR reaction volumes from 5 µl to 5 L
Eliminate spin columns or magnetic beads—Significantly
decrease time and expense while increasing yield
High quality—accurate results
Compared to alternative PCR cleanup methods, ExoSAP-IT Express
reagent provides accurate and consistent results in just 5 minutes.
This unique, highly stable, one-tube solution allows for 100%
recovery of DNA and longer read lengths for greater confidence,
consistency, and accuracy. Treated PCR products produced supe-
rior results when treated with ExoSAP-IT Express reagent (Fig. 1).
ExoSAP-IT Express reagent: PCR cleanup in 5 minutesFastest PCR cleanup method
Our researchers have engineered a heat labile Exonuclease I,
enabling rapid enzyme inactivation in only 1 minute. This
innovation from the original ExoSAP-IT reagent lets scientists
continue their workflow without disruptions. ExoSAP-IT Express
reagent offers faster turnaround times and improved efficiency
of resources while delivering superior sequencing results (Fig. 2).
ExoSAP-IT Express PCR Product Cleanup
Product code Pack size
75001 40 μl 20 reactions
75001 200 μl 100 reactions
75001 1 ml 500 reactions
75001 4 × 1 ml 2,000 reactions
75001 10 ml 5,000 reactions
75001 1 ea 480 reactions × 8-tube strip
Fig. 1. Sequencing results when treated with ExoSAP-IT Express reagent
Sequencing of a 1 kb PCR product treated with ExoSAP-IT Express. ExoSAP-IT Express treatment prior to sequencing eliminates miscalls and improves sequencing scores (numbers and bars above sequence; >60, probability of error ≤0.0001%). Sequence shown is approximately 400 bases from primer binding site.
Conserve PCR samples—simple one-step, 100% recovery
The ExoSAP-IT Express enzymatic cleanup method minimizes
errors by reducing your protocol to a single pipette step, allowing
for automated or manual processing. ExoSAP-IT Express reagent
outperforms the competition by ensuring 100% recovery of all
amplicon sizes, both short and long (Table 1).
Table 1. DNA recovery after purification
PCR size Agencourt AMPure® XP beads ExoSAP-IT reagent
86 bp 10% 100%
103 bp 12% 100%
545 bp 63% 100%
1007 bp 88% 100%
Fig. 2. Compare cleanup methods
Use of ExoSAP-IT Express reagent eliminates spin columns, magnetic beads, sedimentations, filtrations, and gel purifications. With a 5 minute protocol, ExoSAP-IT Express reagent is the fastest and easiest method for PCR cleanup, minimizing pipetting errors or contamination.
HT ExoSAP-IT Fast High-Throughput PCR Product Cleanup is an
alternative formulation of the original ExoSAP-IT reagent specifi-
cally designed for the unique requirements of high-throughput,
automated platforms and multi-channel pipettes. HT ExoSAP-IT
Fast reagent quickly provides accurate and consistent results. This
formulation offers a decreased viscosity and is ideal for automated
liquid handling platforms, with the same convenience and stability
that you have come to expect from ExoSAP-IT reagent.
Also available in a Fast, high-throughput formulation 14 minute cleanup protocol
Stable at 4°C for 1 month and at room temperature for
2 days
Scalable for PCR cleanup from a single tube up to a 384-well
plate PCR reaction
HT ExoSAP-IT Fast High-Throughput PCR Product Cleanup
Product code Pack size
78595 20 rxn 20 reactions
78595 1 ea 480 reactions × 8-tube strip
78595 1 pk5,760 reactions (12 × 8-tube
strips in a tray)
78595 4 pk23,040 reactions (48 × 8-tube
strips in a tray)
78595 1000 rxn 1,000 reactions (2 ml)
78595 5000 rxn 5,000 reactions (10 ml)
78595 1 pl1,920 reactions (96-well
plate, 40 µl per well)
78595 6 pl11,520 reactions (6 × 96-well
plate, 40 µl per well)
Treat 5 μl of PCR product with 2 μl of ExoSAP-IT reagent. Treatment is carried out at 37°C followed by an incubation period at 80°C to completely inactivate both enzymes. Once these contaminants are removed, your PCR products are ready for downstream applications such as sequenc-ing (Sanger/NGS), fragment analysis, SNP analysis, in vitro transcription, or single base extension.
The original ExoSAP-IT formulationConsidered the gold standard in enzymatic PCR cleanup,
ExoSAP-IT PCR Product Cleanup has been cited in over 10,000
publications. Affymetrix is the only manufacturer of USB
ExoSAP-IT reagents, ensuring your lab of product consistency
and integrity.
30 minute cleanup protocol
Recommended for processing 1-96 samples at a time
Best cost per reaction value
Fig. 3. How ExoSAP-IT reagent works
37°C, 4 min for treatment 80°C, 1 min to inactivate
Add ExoSAP-IT®
Express Reagent
* Representative of all ExoSAP-IT formulations. Incubation times differ for ExoSAP-IT Express, original ExoSAP-IT, and HT ExoSAP-IT Fast reagents.
ExoSAP-IT PCR Product Cleanup
Product code Pack size
78250 40 μl 20 reactions
78200 200 μl 100 reactions
78201 1 ml 500 reactions
78202 4 × 1 ml 2,000 reactions
78205 10 ml 5,000 reactions
Overview
ExoSAP-IT reagents are a patented mixture of Exonuclease I
combined with Shrimp Alkaline Phosphatase (SAP) in a specially
formulated buffer that removes excess primers and dNTPs
following a PCR reaction (Fig. 3). Exonuclease I removes residual
single-stranded primers and any extraneous single-stranded DNA
produced during PCR. SAP removes the remaining dNTPs from
the PCR mixture which may interfere with subsequent reactions.
For Research Use Only. Not for use in diagnostic procedures. Affymetrix, USB and ExoSAP-IT are registered trademarks of Affymetrix, Inc. Please see affymetrix.com/trademarks for a complete list of Affymetrix trademarks. All other trademarks are the property of their respective owners. ExoSAP-IT PCR Product Cleanup product line—These products are covered by patents owned by Affymetrix, including US Patent Nos. 6,379,940 and 6,387,634 and corresponding patents in Canada and Japan.
© 2008-2016 Affymetrix, Inc. All rights reserved. Rev 07/15/16P/N USB05033 Rev. 5
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Fax: +65 63957300
Which ExoSAP-IT PCR cleanup reagent is right for you?
ExoSAP-IT® ExpressExoSAP-IT reagentOur original formulation
HT ExoSAP-IT Fast
High-Throughput For automated liquid handlers
Protocol time 5 minutes 30 minutes 14 minutes
Format Single tube 8-tube strips 96-well plate
Single tube Single tube 8-tube strips96-well plate
Throughput level Low to high; Recommended for process-ing any sample size
Low to mid; Recommended for process-ing 1-96 samples at a time
High; Recommended for process-ing ≥ 96 samples at a time
Platform Single- or multi-channel pipette, automated liquid handling platforms
Single-channel pipette Automated liquid handling platforms
Freezes at -20°C No No Yes
Stability -20°C for up to 2 years -20°C for up to 2 years -20°C for up to 2 years;Once thawed, stable at 4°C for 1 month and RT for 2 days
NEW
ExoSAP-IT products offer a unique, one-tube, one-step, enzymatic method for PCR product cleanup.
All ExoSAP-IT reagents provide 100% sample recovery with no loss of PCR products regardless of the fragment size.
This PCR cleanup method removes excess primers and dNTPs and does not interfere with downstream applications.
Achieve superior results with ExoSAP-IT reagent—improve accuracy with higher yields and full PCR product recovery.
Use our comparison chart below to determine which formulation is best for your next experiment.
