HRAS gene mutation detection specificity primer and liquid chip thereof

Abstract

The invention discloses an HRAS gene mutation detection specificity primer and a liquid chip thereof. The liquid chip mainly comprises: an ASPE primer composed of a 5'-terminal tag sequence and 3'-terminal specificity primer sequences focused on target gene mutation sites, wherein the specificity primer sequences comprise SEQ ID NO.16, SEQ ID NO.17, SEQ ID NO.18, SEQ ID NO.19, SEQ ID NO.20 and / or SEQ ID NO.21 focused on a Codon12 site, SEQ ID NO.22, SEQ ID NO.23, SEQ ID NO.24 and / or SEQ ID NO.25 focused on a Codon13 site, and / or SEQ ID NO.26, SEQ ID NO.27, SEQ ID NO.28, SEQ ID NO.29 and / or SEQ ID NO.30 focused on a Codon61 site; a microsphere coated by an anti-tag sequence; and an amplimer. The consistency between the detection result of the detection liquid chip provided by the invention and the detection result of a sequencing method is high to 100%, and the wild-type and mutant parallel detection of a plurality of mutation sites is realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a HRAS gene mutation detection specific primer and a liquid phase chip. Background technique [0002] ras gene participates in the regulation of cell growth and differentiation, and participates in the formation and development of various tumors. There are three characteristic genes related to human tumors in the ras gene family, namely H-ras, K-ras and N-ras, which are located on chromosome 11, 12 and 1 respectively. The activation of ras family oncogenes is the molecular basis for the occurrence and development of some tumors. Among them, point mutations are an important way for the activation of ras family genes, and the point mutations of the 12th, 13th and 61st codons of ras family genes can make them acquire ability to transform cells. Therefore, the detection of ras gene point mutations in tumor cells is of great significance for un...

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.

Again, the above methods all have limitations in detection throughput, and can only detect one mutation type at a time, which cannot meet the needs of practical applications.

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