Specific primers and liquid-phase chip for detection of KITLG gene

Abstract

The invention discloses specific primers and a liquid-phase chip for the single nucleotide polymorphism (SNP) detection of the kit ligand gene (KITLG). The liquid-phase chip mainly comprises allele specific primers (ASPE), each of which consists of a tag sequence at a 5' end and specific primers aiming at mutational sites of target genes at a 3' end, microspheres wrapped by an anti-tag sequence and amplification primers, wherein sequences of the specific primers are one or more from the following pairs of sequences: SEQ ID No. 19 and SEQ ID No. 20, SEQ ID No. 21 and SEQ ID No. 22, SEQ ID No. 23 and SEQ ID No. 24, SEQ ID No. 25 and SEQ ID No. 26, SEQ ID No. 27 and SEQ ID No. 28, SEQ ID No. 29 and SEQ ID No. 30, SEQ ID No. 31 and SEQ ID No. 32, SEQ ID No. 33 and SEQ ID No. 34 and SEQ ID No.35 and SEQ ID No. 36. In the detection liquid-phase chip, the coincidence rate of detection results and a sequencing method is up to 100 percent, so the parallel detection of the wild type and mutanttype of polymorphic loci is realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, and in particular relates to a KITLG gene detection specific primer and a liquid phase chip. Background technique [0002] KITLG (the KIT ligand gene) is the KIT ligand gene located at 12p22. KITLG mutation is a gain-of-function mutation that can increase the amount of melanin in pigment cells, that is, KITLG gene mutation causes familial progressive hyperpigmentation. [0003] At present, there are few reports on the detection and analysis of KITLG gene polymorphisms, and the research methods are mainly direct sequencing and PCR-RFLP analysis. 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 ...

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.

In addition, 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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