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Composite amplification system of 28 short tandem repeats, kit and application thereof

A compound amplification system and short tandem repeat technology, which is applied in recombinant DNA technology, microbial measurement/testing, DNA/RNA fragments, etc., to achieve high individual recognition, short time consumption, and high polymorphic information.

Inactive Publication Date: 2018-03-27
SUZHOU MICROREAD GENETICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the kits currently used are 13 CODIS (Combined DNA Index System) selected by the US FBI: CSF1PO, FGA, TH01, TPOX, vWA, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D21S11, However, in some identifications, more loci are required to provide more information to avoid misjudgment that may occur in some cases

Method used

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  • Composite amplification system of 28 short tandem repeats, kit and application thereof
  • Composite amplification system of 28 short tandem repeats, kit and application thereof
  • Composite amplification system of 28 short tandem repeats, kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Obtaining Individual STR Typing Using the Multiplex Amplification System of 28 Short Tandem Repeats

[0046] 1. Collection of blood samples (blood samples are donated by volunteers)

[0047] 2. DNA extraction

[0048] Genomic DNA was extracted using the Chelex-100 method (refer to "Forensic DNA Protocol". HumanaPress, 1998). Take 0.5-5 μl of anticoagulated whole blood or (1-3mm)*(2-5mm) blood spots in a 500 μl centrifuge tube, Shake and mix Chelex solution to fully suspend Chelex, add 195 μl Chelex-100 (5%) solution and 5 μl proteinase K (20mg / ml) to each tube, shake and mix, keep warm at 56 degrees Celsius for two hours or overnight, take out and shake for 2 minutes, boil water After heating in medium for 10 minutes, centrifuge at 13,000 rpm for 5 minutes, and carefully pipette 150 μl of the supernatant into a new centrifuge tube.

[0049] 3. Reaction system

[0050] Shake and mix each reaction reagent (buffer, primer mix, genomic DNA, etc.) and make a PCR...

Embodiment 2

[0059] Example 2 Using the multiplex amplification system of 28 short tandem repeats for paternity identification

[0060] In this case, a group of triplet families was identified for kinship. Utilize the system of the present invention to identify and obtain each individual type of the triplet family as follows (Table 7):

[0061] Table 7: Individual STR typing in triplet families

[0062]

[0063]

[0064] According to the calculations of mother-child doublet paternity test and triplet paternity test respectively, it is found that the six-color fluorescent multiplex amplification system contains more sites, and the final CPI index is higher than that of the five-color fluorescent multiplex amplification system commonly used in the market today. The CPI index of the system is high. The CPI (Table 8) of the six-color fluorescent composite amplification system is 1.956*10 of the CPI (Table 9) of the five-color fluorescent composite amplification system during the patern...

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PUM

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Abstract

The invention relates to the technical field of biology, and relates to a composite amplification system of 28 short tandem repeats, a kit and an application thereof. The composite amplification system comprises 28 pairs of primers, and can be used for amplifying 28 sites at the same time; and the invention also discloses a primer sequence aiming at the 28 sites. By means of a research of geneticdiversity of a STR locus, 28 sites which contain 22 new CODIS sites, 4 sites with high discrimination power (D4S2366, D6S1043, D11S2368, D14S608) and one gender recognition site Amel, and one Y chromosome insertion-deletion site Y-indel are selected. The sites have higher discrimination power, excluding probability of paternity and polymorphism information content, and other characteristics, so that paternity test and discrimination power system effectiveness can reach a higher level.

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to a system for compound amplification of 28 short tandem repeat sequences of human autosomes. Detection kit, the system can quickly and accurately determine the STR typing of human autosomes, and then provide a basis for forensic individual identification and paternity identification. Background technique [0002] Human genome STR (short tandem repeat sequence) is a relatively stable sequence in DNA inheritance formed by tandem repetition of several bases as the core unit. The distinction between different races, different populations and even different individuals is through the difference in core unit sequence and repeat number, which also constitutes the genetic polymorphism of STR. In the genome, there is an STR site every 15-20kb on average, accounting for 10% of the genome, mostly in non-coding regions and introns, the repeat unit is 2-6bp, the number of repeats is 10-60 times, an...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156C12Q2600/16
Inventor 于在亮杨凡郑玉徐健陈初光
Owner SUZHOU MICROREAD GENETICS
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