High-throughput gene chip for detecting splicing condition of nucleic acid fragment and preparation method of high-throughput gene chip

Abstract

The invention belongs to the field of medical examination and gene chips, and provides a high-throughput gene chip for detecting splicing conditions of nucleic acid fragments and a preparation method of the high-throughput gene chip. The application method of the gene chip comprises the following steps: carrying out reverse transcription on a sample RNA to obtain double-stranded cDNA, carrying out in-vitro transcription to obtain cRNA, adding a fluorescence label, hybridizing the cRNA with a probe on the chip, and carrying out comparative analysis according to the probe position and fluorescence intensity distribution when the chip is designed to obtain the splicing condition of the sample. Compared with the existing whole exon / whole genome detection, the detection cost and the data processing difficulty required by the method are greatly reduced.

Description

technical field [0001] The invention belongs to the field of medical testing and gene chips, in particular to a high-throughput gene chip for detecting the splicing of nucleic acid fragments and a preparation method thereof. Background technique [0002] RNA Microarrays technology / Gene chip technology is a biological detection technology that was developed in the 1980s, matured, promoted and widely used in the 1990s. [0003] Gene chips are used to qualitatively or quantitatively measure nucleic acids present in organisms; they are based on two-dimensional arrays on solid substrates (usually glass slides or silicon films), and the probes used for qualitative and quantitative measurements are composed of several kb (cRNA chips). ) or dozens (oligonucleotides) of specific types of nucleotides. The principle of the gene chip is the complementary pairing between bases and the hybridization between complementary nucleotide single strands. Compared with the traditional Southern ...

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Problems solved by technology

[0009] At present, for the detection of gene mutations, the most commonly used is high-throughput first-generation or second-generation whole-genome sequencing; but this method has two major defects, the first is high cost; Genome sequencing can only measure genes with high expression levels; genes that are usually harmful to humans (disease genes) are often non-wild-type genes, and their expression levels are usually very low; that is, whole-genome sequencing discriminates against genes with low expression levels; and Gene mutations caused by gene splicing are all mutant genes, and whole genome sequencing has no corresponding detection ability for such mutations

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