Method for detecting single base Indel mutation

Abstract

The invention discloses a method for detecting single base Indel mutation, which comprises the following steps of: designing a primer pair of Pw and Pm according to a known existing Indel site in a gene, and detecting and interpreting by performing PCR amplification and agarose gel electrophoresis; extracting the whole genome of a sample to be detected, and respectively performing PCR amplification by taking the whole genome as a template and taking the Pw and the Pm as primers; and isometrically mixing the product obtained by the PCR amplification for agarose gel electrophoresis, and judging whether the Indel of a target gene of the sample to be detected mutates according to the fact that whether a corresponding stripe of the electrophoresis exists. The method can conveniently and accurately detect the Indel mutation of the target gene of the sample to be detected, and has high application value. The Indel mutation has significant practical value on the research of the significant aspects of complex diseases, molecular breeding, genetic diversity evaluation and the like.

Description

Technical field [0001] The field of biotechnology of the present invention relates to the screening and detection of genetic molecular markers, in particular to a method for detecting single-base Indel (single nucleotide insertion / deletion) mutation. Background technique [0002] The genetic information of life is stored in the DNA sequence, and all the DNA of each cell of higher organisms constitutes the organism's genome. Variation in genomic DNA sequence is the basis of species genetic diversity and the original source of variation in biodiversity. Single Nucleotide Variation (SNP) is the most common and basic variation in DNA sequence and becomes the third-generation molecular marker. In the study of human polygenic genetic diseases, SNP analysis has great application potential. For example, SNP markers have been used to find marker genes linked to some important human diseases (diabetes, schizophrenia, cancer, sickle cell anemia, hemophilia, etc.); in crop breeding, import...

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Problems solved by technology

[0004] However, current methods that can detect single nucleotide Indels have certain limitations.

For example: DNA sequence determination is the most accurate method for detecting Indel, but its detection cost is extremely expensive, and requires large-scale instruments such as DNA sequencers and very skilled technicians, so DNA sequence determination is not an application Ideal Indel detection method for actual production; PCR-SSCP experiment process is relatively long, the operation is relatively cumbersome, and there are false negative problems in the experimental process, so it is not an ideal SNP detection method

Restriction fragment length polymorphism (RFLP) is a simple Indel detection method, but its application is very limited and it is not a general method; detection technology platforms such as SNP gene chip, microsphere array technology, mass spectrometer and TaqMan fluorescent probe , which is not very practical for general molecular laboratories

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