Method of capturing genomic target sequence for high-throughput sequencing

Abstract

The invention discloses a method of capturing a genomic target sequence for high-throughput sequencing. The method can sequentially include the following steps of (1) preparing a genomic DNA library;(2) hybridizing with a capture probe library, wherein the capture probe library is composed of n specific probes, n is a natural number, and the specific probes are single-stranded DNA molecules whichare designed and synthesized according to the nucleotide sequence of the target sequence and subjected to locked nucleic acid modification. The capture probe library is composed of 440 probes including the LNA probe-1 to the LNA probe-440; the nucleotide sequences of the LNA probe-1 to the LNA probe-440 are sequentially shown as sequence 1 to sequence 440 in a sequence list. Experiments prove that the capture probe library can capture the corresponding target sequences in a BRCA1 gene and a BRCA2 gene. Therefore, the provided method has important application value.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for capturing genome target sequences for high-throughput sequencing. Background technique [0002] Since the completion of the human genome map, governments of various countries have increased their support for human genome research. Under such a background, gene sequencing technology has developed rapidly, and the gene sequencing market has also expanded rapidly, and has already moved away from the laboratory. into the clinic. Predict your future health status through genetic testing, carry out targeted health care and treatment, and move from passive preventive treatment to proactive health prediction. In the developed countries of the United Kingdom and the United States, genetic testing is as popular as physical examinations. There are more than 6,000 genetic testing items registered by the National Health Service of the United States. The number of genetic testing peo...

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Problems solved by technology

[0004] Part of the cost of gene sequencing comes from the purchase of sequencing equipment, and most of it is consumed on sequencing reagents

At present, domestic gene sequencing reagents mainly rely on imports, and the sequencing reagent market is monopolized by European and American companies, resulting in high prices for sequencing reagents

This has greatly reduced the profit margins of domestic gene sequencing service companies, hindered the promotion and application of sequencing technology in China, and is not conducive to the large-scale industrialization of genetic testing services

[0005] Based on liquid-phase capture high-throughput sequencing technology, the market mainly includes Agilent's whole-exon capture kit, Roche's solid-phase hybridization method and liquid-phase hybridization method. Although the DNA sequences captured by these products are only the whole genome Compared with whole genome sequencing, it has great advantages in terms of cost and time spent in some areas, but the price of each sample is still more than 10,000 yuan, which is beyond the affordability of ordinary people and is not suitable for large-scale screening. Detection of Genetic Diseases

Moreover, the disadvantages of these technologies are the need to synthesize long fragments (150-200bp) of DNA, which are amplified by PCR and transcribed into biotin-modified RNA. The efficiency of probe synthesis is low, the cost is high, the stability of pure RNA probes is poor, and the validity period is short. , the quality is difficult to guarantee

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