High-flux STR sequence core replication number detection method

Abstract

The invention provides a high flux STR sequence core replication number detection method with characteristics of high efficiency and rapidity, which is characterized in that a pair of detection primers are hybridized on a STR sequence cluster amplified on a detection substrate, wherein a fluorescence group is modified on a final primer, then a nucleotide combination is used for extension step by step, simultaneously, the fluorescence signal is detected after each time of extension until the basic group with fluorescence on the final primer is cut by polymerase, then the signal disappears, and then the condition of the fluorescence signal is finally analyzed to obtain the corresponding STR sequence core replication number. According to the invention, an employed biochemical reagent is widely used for the detection platforms of biology chip and high flux sequencing, the signal to noise ratio is high, accuracy for STR detection and resolution to STR heterozygosis can be obviously increased, The method can obtain more convinced result through detection of more STR locus due to high flux characteristic of the biochemical reagent and can detect more samples to realize rapid colony STR detection.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an efficient and rapid high-throughput method for detecting the core repeat number of STR sequences. Background technique [0002] The DNA fingerprints discovered in the 1880s created a variety of means to detect DNA polymorphisms (differences in the DNA structure of different individuals or different populations of organisms), such as RFLP (restriction endonuclease Restriction fragment length polymorphism) analysis, tandem repeat sequence analysis, RAPD (random amplified polymorphic DNA) analysis, etc. Various analysis methods are based on the polymorphism of DNA to produce DNA fingerprints with high individual specificity. Because DNA fingerprints have high variability and stable heredity, and are still inherited in a simple Mendelian way, Became the most attractive genetic marker of the time. [0003] In 1985, Dr. Jefferys first applied DNA fingerprinting technology to forensic ...

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Problems solved by technology

[0006] According to the above-mentioned detection principle, it is not difficult to judge that the current STR detection method has the following shortcomings: (1) The detection reaction is based on the fluorescence capillary electrophoresis method similar to the first generation sequencing technology. Large-scale parallel detection of massive samples; (2) Due to the need to perform fluorescent grouping of STRs according to the length of product fragments, this means that this method has a limit on the number of loci detected, and it is difficult to increase the number of STR sequences to be tested. Further improve the identification rate of biological individuals; (3) there are invalid alleles, which may cause differences in the determination results of some loci in different kits; (4) due to the detection of fragment length, the internal core of STR Single nucleotide polymorphism (SNP) sites in repeats cannot be detected

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