Excision repair cross complementation group 2 (ERCC2) gene mutation detection specific primers and liquid chip kit

Abstract

The invention discloses an excision repair cross complementation group 2 (ERCC2) gene mutation detection liquid chip and specific primers. The liquid chip mainly comprises ASPE primers and each one of the primers comprises a tag sequence at the end 5' and a specific primer sequence at the end 3' aiming at a target gene mutation site. The specific primer sequence comprises sequences shown in the formulas of SEQ ID NO. 9 and SEQ ID NO. 10 aiming at the G934A site, sequences shown in the formulas of SEQ ID NO. 11 and SEQ ID NO. 12 aiming at the T128G site, sequences shown in the formulas of SEQ ID NO. 13 and SEQ ID NO. 14 aiming at the C2133T site, and sequences shown in the formulas of SEQ ID NO. 15 and SEQ ID NO. 16 aiming at the A396C site. The liquid chip also comprises anti-tag sequence-coated microspheres and amplification primers. The detection result of the detection liquid chip and the detection result of the sequencing method have a coincidence rate of 100%. The ERCC2 gene mutation detection liquid chip realizes individual or parallel detection of multiple wild type and mutant type sites.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a specific primer for detecting ERCC2 gene mutation and a liquid phase chip kit. Background technique [0002] ERCC2 (excision repair cross completion group2), located at the 19q13.3 position of chromosome 19, has a total of 23 exons, the full length of the gene is 19.0kb, the full length of the mRNA is 2355bp, and it encodes a protein containing 761 amino acids. The protein encoded by ERCC2 is involved in transcription-coupled nucleotide excision repair, which recognizes large-scale lesions unrelated to repair structures, such as large adducts and thymine dimers, and is a member of the basic transcription factor complex BTF2 A component of / TFIIH. This protein has ATP-dependent DNA unwinding activity and belongs to the RAD3 / XPD helicase subfamily. Loss of function of ERCC2 results in the inability of the basic transcription factor complex...

AI Technical Summary

Problems solved by technology

The PCR-RFLP method is based on the change of the restriction endonuclease recognition site caused by gene mutation, such as the loss of the site or the generation of a new site, a specific fragment is amplified by PCR, and then cut and amplified with a restriction endonuclease. Amplify the product and observe the size of the fragment by electrophoresis. This method is used to detect gene mutations with altered restriction sites, and can directly determine the genotype. However, this method cannot be used for the detection of gene mutations without new restriction sites.

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry is a soft ionization technology that has powerful and mature functions in the detection of protein and other biological macromolecules. However, in the field of nucleic acid detection, due to the particularity of nucleic acid molecules, detection is subject to certain limit

Although immunohistochemistry has the advantages of specificity, strong sensitivity, and simple operation, there is no uniform standard for the use of antibodies, operating procedures, and result judgments, which affects its clinical promotion and application.

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