VKORC1 gene polymorphism detection genotyping kit based on AllGlo probe and genotyping method thereof

Abstract

The invention relates to single nucleotide polymorphism, in particular to a VKORC1 gene polymorphism detection genotyping kit based on an AllGlo probe and a genotyping method thereof. The kit comprises a real-time fluorescence quantification PCR (polymerase chain reaction) reagent, positive control and negative control, and VKORC1(1173C>T) is the SNP (single nucleotide polymorphism) locus, provided by NCBI(National Center of Biotechnology Information), of human chromosome 16 in the VKORC1 gene. The genotyping method includes the steps that DNA in an EDTA anticoagulant whole blood sample is extracted by adopting a conventional method; the real-time fluorescence quantification PCR reagent provided by the VKORC1 gene polymorphism detection genotyping kit based on the AllGlo probe is used for conducting real-time fluorescence quantification PCR amplification on DNA; the SNP locus VKORC1(1173C>T) of the human VKORC1 gene is genotyped according to detected fluorescence signals.

Description

technical field [0001] The invention relates to single nucleotide polymorphism (SNP), in particular to a VKORC1 gene polymorphism detection and typing kit and typing method based on AllGlo probe. Background technique [0002] Single nucleotide polymorphism (singlenucleotide polymorphism, SNP) mainly refers to the DNA sequence polymorphism caused by the variation of a single nucleotide at the genome level. It is the most common type of heritable variation in humans. SNPs widely exist in the human genome, with an average of 1 in every 500-1000 base pairs, and it is estimated that the total number can reach 3 million or more. As a third-generation genetic marker, SNP is closely related to many phenotypic differences, drug susceptibility and disease susceptibility. The large number of SNPs in the genome makes it a powerful tool and plays a very important role in disease localization and cloning, drug design and testing, and basic biological research. [0003] Individualized d...

AI Technical Summary

Problems solved by technology

Among them, the direct sequencing method is currently the most intuitive and relatively accurate SNP typing method, but its steps are many and scattered, the cost is high, the workload is heavy, the cycle is long, and the price is expensive, so it is not suitable for large samples and multiple loci. Detection; Restriction Fragment Length Polymorphism (RFLP), as the earliest DNA labeling technology, has low requirements for instruments, and only needs a PCR instrument and an electrophoresis instrument to carry out the experiment, but its experimental operation is cumbersome and the detection cycle is long , not suitable for large sample detection; Tagman fluorescent probe method has good accuracy, but its design and synthesis procedures are complicated, and it is not suitable for typing of multiple loci and a small number of samples, especially loci with low frequencies; amplification block mutation The system (ARMS method) is fast and simple, but it cannot be operated in a closed tube, and it is difficult to achieve high-throughput and automation for gene polymorphism analysis; high-resolution melting curve analysis (HRM) has the advantages of simplicity and speed, but the method is specific Insufficient, few instrument choices

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