KCNQ1 (potassium voltage-gated channel, KQT-like subfamily, member 1), KCNE2 (potassium voltage-gated channel, Isk-like family, member 2) and KCNN3 (calcium-activated potassium (SK) channels) gene SNP (Single Nucleotide Polymorphism) detection specific primer and liquid-phase chip

Abstract

The invention provides a KCNQ1 (potassium voltage-gated channel, KQT-like subfamily, member 1), KCNE2 (potassium voltage-gated channel, Isk-like family, member 2) and KCNN3 (calcium-activated potassium (SK) channels) gene SNP (Single Nucleotide Polymorphism) detection specific primer and liquid-phase chip. The liquid-phase chip comprises an ASPE (Allele Specific Primer Extension) primer, microspheres and an amplification primer, wherein the ASPE primer consists of a tag sequence at 5' end and specific primers aiming at target gene mutation at 3' end, wherein the specific primers are SEQ ID NO. 9 and SEQ ID NO. 10 aiming at a KCNQ1 gene A184G SNP site, SEQ ID NO. 11 and SEQ ID NO. 12 aiming at a KCNQ1 gene A70C SNP site, SEQ ID NO. 13 and SEQ ID NO. 14 aiming at a KCNE2 gene G166A SNP site, and / or SEQ ID NO. 15 and SEQ ID NO. 16 aiming at a G107A gene G166A SNP site; and the microspheres are coated by anti-tag sequence. The matching ratio of the detection result of the KCNQ1, KCNE2 and KCNN3 gene SNP detection liquid-phase chip provided by the invention and that of a sequencing method reaches up to 100 percent.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a KCNQ1, KCNE2 and KCNN3 gene SNP detection specific primer and a liquid phase chip. Background technique [0002] The KQT-like subfamily No. 1 potassium channel gene (potassium voltage-gated channel, KQT-like subfamily, member1, KCNQ1) is located at 11p15.5 of the human genome, about 400kb, and consists of 17 exons. KCNQ1 is a voltage-dependent potassium channel expressed in many tissues, including cochlea, epithelial tissue and cardiomyocytes. Its physiological function is to transport potassium ions out of cells and maintain the ion concentration required for normal physiology. Isk-related family No. 2 potassium channel gene (potassium voltage-gated channel, Isk-related family, member 2, KNCE2) is located at 21q22.12 of the human genome, with more than 35.73 million base pairs, and its main physiological function is to maintain cardiomyoc...

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. The size of the fragment is observed by electrophoresis. This method is used to detect gene mutations with altered restriction sites, and can directly determine the genotype, but this method cannot be used for the detection of gene mutations without new restriction sites; while the traditional Solid-phase chips are expensive, and the sensitivity is not high, and the reproducibility of the test results is poor

Thirdly, both the detection method based on PCR technology (PCR-RFLP) and the allelic difference analysis method based on TaqMan technology have limitations in the detection throughput, and only one mutation can be detected at a time, which cannot meet the needs of practical applications.

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