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Brief Introduction of NC22 Plex STR Detection Kit
1. Five-color fluorescence detection platform, which can carry out multiple amplification and detection on 20 supplemental loci, 1 CODIS locus and 1 sex locus specified in the Standard of Judicial Identification;
2. Super high resolution, sensitivity and anti-inhibitor ability;
3. Rapid amplification, the amplification time is less than 1.5 hours.
4. Combined with our 21 Plex STR STR Detection kit, the number of detected loci can reach 40;
5. Meet the "Code for the Implementation of the Identification of Full Biological Syndrome" published by the Judicial Administration of Justice of the Ministry of Justice of PRC in 2014, it recommends to increase the number of detected loci from 19 to 29 or 39;
6.
7. Can be used in forensic analysis, kinship detection and scientific research for human genetic identification.
Brief Introduction of NC22 Plex STR Detection Kit
1.1 Features of NC22 Plex
Brief Introduction of NC22 Plex STR Detection Kit
1.2 Loci Arrangement Diagram of NC22 Plex
1.3 Analysis Software Loci Information Table of NC22 Plex GeneMapper ID-X
Brief Introduction of NC22 Plex STR Detection Kit
1.4 Electrophoretic Diagram of NC22 Plex Allelic LadderNC22 Plex Allelic Ladder contains 261 allelic genes
Brief Introduction of NC22 Plex STR Detection Kit
Component
PCR Amplification Kit
2.5× PCR Mix buffer containing anti-inhibition components such as MgCl2
NC22 Plex Primers primer mixtures for amplifying 22 loci
PCR Grade Water deionized ultra pure water
Control DNA 9947A(0.5ng/µL) standard positive control
Taq DNA Polymerase hot-start enzyme
PCR Detection KitNC22 Plex Allelic Ladder standard reference material for allele
genotyping
Orange-500 molecular weight internal labels with fluorescent markers
Matrix Kit 5 Dye Matrix used for fluorescence correction
Components of NC22 Plex STR Detection Kit
2.1 NC22 Plex STR Fluorescence Detection Kit (25µL×100 Reactions/Kit)
Component PCR Amplification Kit
2.5× PCR Mix 1000× 1
NC22 Plex Primers 500× 1
PCR Grade Water 1000× 1
Control DNA 9947A(0.5ng/µL) 20× 1
Taq DNA Polymerase 100× 1
Components of NC22 Plex STR Detection Kit
Component PCR Detection Kit
NC22 Plex Allelic Ladder 20× 1
Orange-500 120× 1
Component Matrix Kit
5 Dye Matrix 50× 1
2.2 Components and Contents of NC22 Plex STR Detection Kit
95℃
3min94℃
15s60℃
1min
60℃
30min
4℃
∞30 Cycles
Component 25µL System(µL)
2.5× PCR Mix 10.0
NC22 Plex Primers 5.0
PCR Grade Water 8.0
Control DNA 9947(0.5ng/µL) 1.0
Taq DNA Polymerase 1.0
Total Volume 25.0
Components of NC22 Plex STR Detection Kit
2.3 Reaction System and Procedures of NC22 Plex STR STR Detection Kit
Amplification Results of NC22 Plex STR Detection Kit
3.1 Amplification Results of Positive Control 9947A on 3130xl
Amplification Results of NC22 Plex STR Detection Kit
3.2 Amplification Results of Blood on FTA on 3130xl
Amplification Results of NC22 Plex STR Detection Kit
3.3 Amplification Results of Blood on Filter Paper on 3130xl
Amplification Results of NC22 Plex STR Detection Kit
3.4 Amplification Results of Saliva on FTA on 3130xl
4.1 Sensitivity Test
Performance Measurement of NC22 Plex STR Detection Kit
Ø 4.1.1 Content of Extracted DNA • 1ng, 500pg, 250pg, 125pg, 62.5pg, 31.25pg • Repeat the experiment 3 times for all samples
Ø 4.1.2 Number of Direct Amplification Samples FTA Blood Card • 1, 2 or 3 slices - 1.2mm aperture • Repeat the experiment 3 times for all samples FTA Saliva Card • 1, 2 or 3 slices - 1.2mm aperture • Repeat the experiment 3 times for all samples
Performance Measurement of NC22 Plex STR Detection Kit
4.1.1 Extracted DNA
• 30 cycles• 25μL amplification system• 3130xl (3kV,10sec sampling time)• Baseline threshold 50RFU
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
1ng 500pg 250pg 125pg 62.5pg 31.25pg
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DNA Amount
Performance Measurement of NC22 Plex STR Detection Kit
4.1.1 Extracted DNA
Performance Measurement of NC22 Plex STR Detection Kit
4.1.2 Number of Direct Amplification Samples FTA Blood Card & FTA Saliva Card
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
1 slice 2 slices 3 slices
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Number of 1.2mm punches
blood on FTAbuccal on FTA
• 30 cycles• 25μL amplification system• 3130xl (3kV,10sec sampling time)• Baseline threshold 50RFU
Performance Measurement of NC22 Plex STR Detection Kit
4.1.2 Number of Direct Amplification Samples FTA Blood Card & FTA Saliva Card
Ø 4.2.1 Mix Concentration • 0.75 x, 1 x, 1.25 x
Ø 4.2.2 Primer Concentration • 0.75 x, 1 x, 1.25 x
Ø 4.2.3 Annealing Temperature • 55℃, 57 ℃, 59 ℃, 61 ℃
Ø 4.2.4 Cycles Number • 28, 29, 30, 31
Ø 4.2.5 Reaction Volume • 25µl and 10µl
Performance Measurement of NC22 Plex STR Detection Kit
4.2 Tolerance Test
4.2.1 2 Concentrations of Mix and Primer
Performance Measurement of NC22 Plex STR Detection Kit
• 30 cycles• 25μL amplification system• 3130xl (3kV,10sec sampling time)• Baseline threshold 50RFU
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
0.75X 1X 1.25X 0.75X 1X 1.25X
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Mix Concentration
blood on FTAbuccal on FTA
4.2.1 Mix Concentration
Performance Measurement of NC22 Plex STR Detection Kit
4.2.2 Primer Concentration
Performance Measurement of NC22 Plex STR Detection Kit
• 30 cycles• 25μL amplification system• 3130xl (3kV,10sec sampling time)• Baseline threshold 50RFU
4.2.3 Annealing Temperature
Performance Measurement of NC22 Plex STR Detection Kit
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
56℃ 58℃ 60℃ 61℃
Perc
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Annealing Temperature
blood on FTAbuccal on FTA
4.2.3 Annealing Temperature — FTA Blood Card
Performance Measurement of NC22 Plex STR Detection Kit
4.2.3 Annealing Temperature — FTA Saliva Card
Performance Measurement of NC22 Plex STR Detection Kit
4.2.4 Cycles Number — 25 μL
Performance Measurement of NC22 Plex STR Detection Kit
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
28 29 30 31
Perc
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Cycles Numbers (25uL)
blood on FTAbuccal on FTA提取DNA
4.2.4 Cycles Number — 10 μL
Performance Measurement of NC22 Plex STR Detection Kit
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
28 29 30 31
Perc
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f Loc
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Cycles Numbers (10uL)
blood on FTAbuccal on FTA提取DNA
4.2.4 Reaction Volume
Performance Measurement of NC22 Plex STR Detection Kit
Three kinds of inhibitors:
Ø 4.3.1 Heme • 0.1, 0.2, 0.3 and 0.4mM
Ø 4.3.2 Hemoglobin • 80, 100, 200 and 300µM
Ø 4.3.3 Humic Acid • 10, 30, 50 and 100ng/µL
4.3 Inhibition Test
Performance Measurement of NC22 Plex STR Detection Kit
• 30 cycles • 25μL amplification system• 3130xl (3kV,10sec sampling time)• Baseline threshold 50RFU
4.3 Inhibition Test
Performance Measurement of NC22 Plex STR Detection Kit
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1
0.1mM
0.2mM
0.3mM
0.4mM
60μM
100μ
M20
0μM
300μ
M
10ng
/μL
30ng
/μL
50ng
/μL
100n
g/μL
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Hemoglobin
4.3.1 Heme
Performance Measurement of NC22 Plex STR Detection Kit
4.3.2 Hemoglobin
Performance Measurement of NC22 Plex STR Detection Kit
4.3.3 Humic Acid
Performance Measurement of NC22 Plex STR Detection Kit
1. Fast: Optimized components of PCR Reaction System that can shorten the PCR amplification time to less than 1.5 hours.
2. High resolution: Single tube reaction can get the correct typing of 22 loci at one time, which performs better than the other similar products in the market.
3. DNA extraction-free technology: Samples can be used for PCR amplification directly without extraction.
4. Higher rate of individual recognition and non-parent exclusion, showing excellent performance in criminal investigation, judicial identification and paternity test no matter NC22 Plex is used exclusively or jointly with other kits.
5. The product is fully domestically made, which improves both the independent innovation ability of the core technology of China's DNA detection reagents and the security of national DNA detection industry.
4. Summary
Summary of NC22 Plex STR Detection Kit
Comparison of Autosomal Supplementary Kits between Different Manufacturers
1. Loci Selected by Different Manufacturers
Comparison of Autosomal Supplementary Kits between Different Manufacturers
2. Comparison of Selected Loci