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Fluorescent PCR based method for detecting polymorphism of TYMS gene

A technology of gene polymorphism and fluorescence, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of PCR efficiency reduction, primer extension block, etc., and achieve improved sensitivity, high site specificity, and guaranteed The effect of accuracy

Active Publication Date: 2014-04-30
SHANXI LIFEGEN
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Problems solved by technology

The principle is that the primer extension guided by DNA polymerase starts from the 3' end of the primer in the PCR process, and the degree of complementarity between the base at the 3' end of the primer and the template strongly affects the recognition of the polymerase and the progress of the PCR reaction: if this If the base and the template are normally complementary (A-T, G-C), the primer can be extended without interruption, and the PCR can be efficiently performed to obtain a complete product; on the contrary, if the base is not normally paired with the template, the extension of the primer is blocked. PCR efficiency is greatly reduced

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  • Fluorescent PCR based method for detecting polymorphism of TYMS gene
  • Fluorescent PCR based method for detecting polymorphism of TYMS gene
  • Fluorescent PCR based method for detecting polymorphism of TYMS gene

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Embodiment Construction

[0032] The invention is a technology for detecting TYMS gene 1494del6 polymorphism by fluorescent allele-specific PCR, which can be applied to the analysis of TYMS gene 1494del6 polymorphism in vitro.

[0033] According to the two alleles corresponding to the TYMS gene 1494del6 polymorphism, the present invention designs allele-specific primers respectively, and combines SYBR green technology to accurately determine the polymorphism of the TYMS gene 1494del6 in the sample through the amplification curve after the fluorescent PCR is completed. Specific genotypes of morphological loci. Include the following steps:

[0034] (1) Design of allele-specific primers:

[0035] Design allele-specific primers for the two alleles corresponding to the TYMS gene 1494del6 polymorphism as upstream primers, the sequences are as follows:

[0036] +6bp:5'-GTAGAGTGTGGTTATGAACTTTAA-3'

[0037] -6bp:5'-GTAGAGTGTGGTTATGAACTTTGT-3'

[0038] Design a common downstream primer for the two alleles, t...

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Abstract

The invention provides a rapid and accurate fluorescent PCR based method for detecting 1494del6 polymorphism of a TYMS gene. Aiming at two alleles corresponding to 1494del6 polymorphism of the TYMS gene, the method respectively designs specific primers of the alleles, and an SYBR green technology is combined, so that after the fluorescent PCR, specific genotype of 1494del6 polymorphism locus of the sample can be accurately determined according to an amplification curve. The specific primers designed by the invention have the advanategs of high site specificity, high detection sensitivity and material and time saving.

Description

technical field [0001] The invention relates to a method for detecting polymorphism of TYMS gene. Background technique [0002] Thymidylate Synthase (TS) is the rate-limiting enzyme for pyrimidine nucleotide synthesis, encoded by the TYMS gene. TS catalyzes the conversion of deoxyguanylate (dUMP) to deoxythymidylate (dTMP), an essential step in the DNA synthesis process. TYMS gene 3' non-translated region (untranslated region, UTR) has a 6bp deletion (1494del6) gene polymorphism. [0003] The present invention is to detect the 6bp deletion (1494del6) gene polymorphism (expressed as +6bp / +6bp, +6bp / -6bp and -6bp / -6bp three) occurring in the 3' non-coding region (untranslated region, UTR) of the TYMS gene genotype). [0004] The traditional techniques for detecting TYMS gene 1494del6 polymorphism mainly include direct sequencing and PCR-gel electrophoresis. These two methods are low in cost and high in accuracy, but both require PCR post-processing steps, which are complic...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6888C12Q2600/156C12Q2563/107C12Q2561/113
Inventor 戴鹏高陈超周文静
Owner SHANXI LIFEGEN
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