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Genotyping detection kit for 20 str loci on human chromosomes 13, 18 and 21

A D13S1817, locus technology, applied in the field of genotyping detection kits, can solve problems such as application troubles, achieve the effects of good genotyping map, improve work efficiency, and save costs

Active Publication Date: 2017-08-29
上海春夏正像生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this characteristic of the STR locus has caused problems in its application in the identification of kinship

Method used

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  • Genotyping detection kit for 20 str loci on human chromosomes 13, 18 and 21
  • Genotyping detection kit for 20 str loci on human chromosomes 13, 18 and 21
  • Genotyping detection kit for 20 str loci on human chromosomes 13, 18 and 21

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Preparation of genomic DNA from human blood samples

[0038] Test samples were donated by volunteers with informed consent. According to medical routine, 1 mL of peripheral venous blood was collected and anticoagulated with EDTA. Genomic DNA was extracted using the Human Peripheral Blood Genome Extraction Kit from QIAGEN, with an elution volume of 100 microliters, quantified using a UV spectrometer, and the concentration of genomic DNA was diluted to 1 ng / μL.

Embodiment 2

[0039] Embodiment 2: PCR system preparation

[0040] Prepare the PCR reaction system according to the following system (the total reaction system is 20 μL):

[0041] 2×PCR reaction master mix 10μL

[0042] Primer mix 2 μL

[0043] Sample DNA template 1 μL

[0044] wxya 2 O 7μL

[0045] Take out the primer composition container and the PCR reaction mother solution container from the kit. According to the total number of reactions multiplied by the individual reaction requirements in the above reaction system, the required amount of PCR reaction mother solution, primer composition amount, PCR reaction auxiliary liquid amount and ddH 2 O amount, mix the above reagents evenly in a 1.5mL EP tube, divide and number each PCR reaction tube 19 μL, then add 1 μL of the sample DNA template according to the sample number, and mix again.

Embodiment 3

[0046] Embodiment 3: PCR reaction

[0047] ABI 9700 PCR instrument was used for PCR reaction.

[0048] The PCR conditions are as follows: 95°C for 10 minutes after starting, 95°C for 30 seconds, 60°C for 60 seconds, 72°C for 60 seconds, a total of 30 cycles, then 72°C for 60 minutes, and then 4°C for incubation.

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Abstract

The invention belongs to the technical filed of human nucleic acid in-vitro detection, and more specifically relates to a gene typing detection kit for human 13,18 and 21 chromosome 20 STR locus. According to the invention, a composite PCR amplification system for gene typing on 20 STR locus distributed on human 13, 18 and 21 chromosome is designed, and wherein a PCR amplification primer group is SEQ ID No.1-SEQ ID No. 40. The gene typing detection kit comprises a primer combination container and a PCR reaction mother liquor container; and the primer combination container comprises primer SEQ ID No.1-SEQ ID No. 40 composition storage liquid. In the invention, single tube amplification on 20 STR locus is carried out, STR locus in each fluorescence channel and different fluorescence channels have equal amplification, a typing graph is good; cost, manpower and time can be obviously saved, and work efficiency is increased.

Description

technical field [0001] The invention belongs to the technical field of in vitro detection of human nucleic acid, in particular to the detection of the genotype of the highly polymorphic STR loci in human genome DNA, and in particular to a method for detecting human No. 13, 18 and 21 by multiple polymerase chain reaction Kit for genotyping detection of 20 STR loci on chromosomes. Background technique [0002] Short tandem repeats (Short tandem repeats, STR) are a type of DNA tandem repeats that widely exist in the human genome. The core repeat unit is usually 2-6 bases, and it is the most polymorphic genetic sequence in the human genome. Markers, the polymorphism of which is mainly derived from the difference in the number of repeats of the core repeat unit among individuals, and the repeat number of the core repeat unit follows the Mendelian law of inheritance in the genetic process. Because alleles of the same STR locus (STR locus alleles are usually named according to the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/156C12Q2600/16
Inventor 赵书民周巍龚虎涛
Owner 上海春夏正像生物科技有限公司
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