PCR-Sort-PCR library enriching method applicable to high-throughput sequencing

Abstract

The invention discloses a PCR-Sort-PCR library enriching method applicable to high-throughput sequencing. The method comprises the following steps: (1) in a library constructing process, purifying a joint connecting product, and beginning the first turn of PCR reaction; (2) after the first turn of PCR, purifying obtained nucleic acid or conducting fragment separation; and (3) conducting the second turn of PCR reaction by taking a purified product of the first turn of PCR or a product of the fragment separation as a template. With the development of the technology, more and more serious sample problems appear in the high-throughput sequencing, mainly manifested in that the total amount of samples becomes less and less and complexity becomes higher and higher. In relative terms, the amount of the samples is the most limiting factor. It is especially important to guarantee the smooth completion of the high-throughput sequencing on the basis of the circumstance that it is impossible to increase the total amount of the samples; with the application of the library enriching technology of the parent, the problem that the total amount of the samples is insufficient in a library constructing process is solved, an obtained library is relatively concentrated in fragments and relatively high in yield, and meanwhile sample abundance is guaranteed.

Description

technical field [0001] The invention relates to the technical field of high-throughput sequencing of genes, in particular to a PCR-Sort-PCR (PSP) library enrichment technology suitable for high-throughput library construction of ultra-low starting nucleic acid templates. Background technique [0002] In second-generation and third-generation sequencing technologies, constructing high-quality sequencing libraries is a key factor for high-throughput sequencing. The quality of the sequencing library directly determines the quality of the data produced by the sequencing, which in turn has a great relationship with the data analysis. With the rapid development of high-throughput sequencing technology, major reagent companies, high-throughput sequencing has been greatly applied in various fields such as zoology, botany, microbiology, agriculture, and medicine. High-throughput sequencing library construction methods are gradually being developed, matured, and applied; the types of...

AI Technical Summary

Problems solved by technology

High-throughput sequencing library construction methods are gradually being developed, matured, and applied; the types of samples faced are becoming more and more complex, such as single cells, cfDNA, etc. It is not easy to obtain such samples, and it is difficult to meet the needs of high-throughput sequencing libraries. build requirements

In this case, a variety of library construction methods have been developed. For example, for single-cell samples, there are existing single-cell transcript library construction methods and kits, and single-cell genome library construction methods and kits. Specific library construction methods and kits for the development of input samples

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