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Method for preparing human gene STR (short tandem repeat) sequence allelic ladder by using site-directed mutagenesis technology

A typing standard, short tandem repeat technology, applied in the field of biomedicine to achieve the effect of improving efficiency

Inactive Publication Date: 2016-05-25
THE FIRST RES INST OF MIN OF PUBLIC SECURITY +1
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Problems solved by technology

Replication slippage, unequal exchange of sister chromosomes and genetic recombination are all mechanisms that can lead to changes in the number of repeat units, but the current study proves that the change in the number of STR repeats caused by replication slippage is the main mechanism

Method used

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  • Method for preparing human gene STR (short tandem repeat) sequence allelic ladder by using site-directed mutagenesis technology
  • Method for preparing human gene STR (short tandem repeat) sequence allelic ladder by using site-directed mutagenesis technology
  • Method for preparing human gene STR (short tandem repeat) sequence allelic ladder by using site-directed mutagenesis technology

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Embodiment Construction

[0037] The present invention will be further described below in conjunction with the accompanying drawings. It should be noted that this embodiment is based on the technical solution, and provides detailed implementation and specific operation process, but the protection scope of the present invention is not limited to the present invention. Example.

[0038] like figure 1 As shown, a method for preparing a human gene short tandem repeat sequence allelic typing standard by site-directed mutagenesis technique comprises the following steps:

[0039] S1 Design amplification primers according to the sequence of the target STR locus, and use any individual human DNA as a template to amplify;

[0040] S2 Cloning the obtained amplified product into a plasmid vector, then transforming it into DH5α competent cells, and plating for culture;

[0041] S3 picks a single clone for sequencing, compares the sequencing results with the original sequence, and saves the recombinant plasmid wit...

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Abstract

The invention discloses a method for preparing a human gene STR (short tandem repeat) sequence allelic ladder by using the site-directed mutagenesis technology. The typing of the known STR locus is disclosed; in the invention, homologous arm primers are designed according to different types of the STR locus; different types of a locus are obtained by a site-directed mutagenesis method, and then the obtained different locus types are cloned into plasmids; and therefore, the work of early sample screening is saved, and the preparation efficiency of an STR ladder is improved.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for preparing a human gene short tandem repeat sequence allele typing standard by using site-directed mutagenesis technology. Background technique [0002] Allelic typing standards refer to the common allelic mixture of a certain STR locus in the population, which provides a reference for the size of DNA fragments for each allele. Allelic typing standards are very important for the accurate typing of STR loci. They can correct fragments of different sizes detected for the same allele in different laboratories due to different instruments and testing conditions. standard. [0003] The repeat sequence with repeat unit length of 2-6bp is called short tandem repeat (shorttandem repeat, STR). The repeated sequences of STR loci are arranged in tandem, and the gene amplification fragments are generally below 400bp. Currently, STRs are the most commonly used class of gene...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12N15/70C12Q1/68
CPCC12Q1/6806C12Q2531/113
Inventor 姜伯玮荣海博管桦俞丽娟聂艳钗赵颖金川张涛
Owner THE FIRST RES INST OF MIN OF PUBLIC SECURITY
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