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Multiple PCR (polymerase chain reaction) primer combination and detection method used for human paternity test

A paternity test and primer combination technology, applied in the field of human paternity test detection systems, can solve problems such as incapacity

Inactive Publication Date: 2013-06-26
上海邃志生物科技股份有限公司
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AI Technical Summary

Problems solved by technology

However, when the needs of identification are extended to more distant relatives, such as when a disaster victim has only one cousin who can provide the samples required for identification, such STR identification is somewhat inadequate

Method used

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  • Multiple PCR (polymerase chain reaction) primer combination and detection method used for human paternity test
  • Multiple PCR (polymerase chain reaction) primer combination and detection method used for human paternity test
  • Multiple PCR (polymerase chain reaction) primer combination and detection method used for human paternity test

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0027] Example: Using time-of-flight mass spectrometry to perform SNP polymorphism typing and parent-child inference for the population.

[0028] A) Extract genomic DNA

[0029] 1. Blood DNA extraction method

[0030] The sample for this test is whole blood added with anticoagulant (EDTA or sodium citrate) and stored in cold storage. The specific steps of DNA extraction are as follows:

[0031] 1) Take 200μl whole blood and add 1ml H 2 O, mix upside down, centrifuge at 3000rpm for 2min, discard the supernatant, repeat 2-3 times until the red color disappears.

[0032] 2) Add 400μl of 1×cell lysate, 5μl of proteinase K, and water bath at 56℃ for 30min.

[0033] 3) Take out the sample in an ice water bath for 1 min, add 150μl of 6mol / L NaCl, and shake vigorously for 15-20 seconds. Centrifuge at 13000 rpm for 10 minutes.

[0034] 4) Pipette the supernatant into a new centrifuge tube, add 1.1ml of absolute ethanol, turn upside down 10 times until flocculent DNA precipitation appears, leave i...

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Abstract

The invention relates to a multiple PCR (polymerase chain reaction) primer combination used for a human paternity test. Eighty SNP (single nucleotide polymorphellosm) genetic markers can be detected by virtue of the primer combination. Nucleotide sequences of primers at middle and upper reaches in the primer combination are respectively shown in SEQ ID NO.1-80; nucleotide sequences of primers at a lower reach in the primer combination are respectively shown in SEQ ID NO.81-160; and nucleotide sequences of single-base extension primers are sequentially shown in SEQ ID NO.161-240. According to the multiple PCR primer combination, the human paternity test is carried out by preferably utilizing a flight time spectral method, a result is accurate, a method is rapid and simple, and flux is high, so that the multiple PCR primer combination has a good application prospect.

Description

Technical field [0001] The present invention relates to a detection system for human paternity testing by using a set of multiple PCR primer combinations to detect 80 SNP genetic markers, and specifically relates to a detection system of 80 upstream primers, 80 downstream primers and 80 single-base extension primers Nucleotide sequence and detection method. Background technique [0002] The modern method of kinship identification is DNA analysis, which uses genetic markers on human chromosomes to identify whether there is a blood relationship between two individuals. The probability of sharing the same genetic markers between individuals who are related is greater than that of individuals who are not related. At present, the genetic marker used for paternity testing is the second-generation genetic marker—short tandem repeat (STR), which has a wide distribution, is easy to detect, has a large amount of information, is highly polymorphic, and follows Mendelian codominant inherita...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 张宇清王健
Owner 上海邃志生物科技股份有限公司
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