RB1 gene mutation detection specific primer and liquid chip

Abstract

The invention discloses an RB1 gene mutation detection specific primer and a liquid chip. The liquid chip mainly comprises ASPE primers consisting of tag sequences at a 5' end and specific primer sequences aiming at target gene mutation sites at a 3' end, microspheres coated with anti-tag sequences and an amplification primer, wherein the specific primer sequences are as follows: SEQ ID NO.7 and SEQ ID NO.8 which aim at a C1654T site, SEQ ID NO.9 and SEQ ID NO.10 which aim at a C1666T site, and / or SEQ ID NO.11 and SEQ ID NO.12 which aim at a C1981T site. The detection liquid chip provided by the invention has the advantages that the matching rate between the detection result and a sequencing method is up to 100%, and the wild-type and mutant parallel detection of multiple mutant sites can be realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a specific primer for RB1 gene mutation detection and a liquid phase chip. Background technique [0002] Retinoblastoma 1 (RB1) is the first human tumor suppressor gene isolated and cloned. Because this gene is closely related to the occurrence of retinoblastoma, it was named retinoblastoma gene. The RB1 gene is a negative regulator of the cell cycle, which regulates the expression of genes required for cell proliferation and differentiation by combining with transcription factors, thereby maintaining the balance of cell growth and development. Therefore, the function of this gene is related to cell cycle, cell senescence, cell apoptosis, cell differentiation and growth inhibition, etc. [0003] At present, there are few methods for detecting and analyzing RB1 gene mutations, mainly including direct sequencing and single-strand conformatio...

AI Technical Summary

Problems solved by technology

The SSCP analysis method is mainly to separate single-stranded DNA molecules with different spatial structures through non-denaturing polyacrylamide gel electrophoresis (PAGE), and analyze the results by autoradiography, silver staining or ethidium bromide color development. Comparing the band mobility with the normal control, it is determined that the chain conformation has changed, and then it is inferred that there is a base mutation in the DNA fragment. This method is simple and fast; however, the SSCP analysis method cannot determine the position and type of the base mutation. Electrophoretic conditions are strictly required. In addition, since SSCP achieves electrophoretic separation based on the change of the three-dimensional conformation of single-stranded DNA molecules caused by point mutations, it may occur that when point mutations at certain positions do not affect the three-dimensional conformation of single-stranded DNA molecules. When it works or the effect is small, coupled with the influence of other conditions, polyacrylamide gel electrophoresis cannot distinguish and cause missed detection

Again, 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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