Ultramicro nucleic acid sample library building method applied to NGS platform

Abstract

The invention discloses an ultramicro nucleic acid sample library building method applied to an NGS platform. The ultramicro nucleic acid sample library building method comprises the following steps: adding nucleic acid repair enzyme into an ultramicro nucleic acid sample for nucleic acid repair before DNA extraction of the ultramicro nucleic acid sample; then carrying out DNA extraction on the ultramicro nucleic acid sample containing the nucleic acid repair enzyme; and carrying out library construction on the extracted DNA to obtain a sequencing library of the ultramicro nucleic acid sample. According to the ultramicro nucleic acid sample library building method, nuclease repair is carried out on sample DNA before DNA extraction; in addition, nucleic acid repair enzyme is added in the DNA extraction process; DNA damage can be effectively repaired, reaction false positive is reduced, DNA extraction quality is improved, and the constructed library can be better suitable for NGS sequencing. According to the library building method, the problems that the DNA extracted from the FFPE sample is poor in quality and low in total quantity, and the library cannot be effectively built are solved.

Description

technical field [0001] This application relates to the technical field of high-throughput sequencing, in particular to a method for building a library of ultra-trace nucleic acid samples applied to an NGS platform. Background technique [0002] Next-generation sequencing (NGS) systems are well known for their ability to simultaneously perform a large number of parallel sequencing reactions. Although different next-generation sequencing platforms have many differences in technical details, libraries need to be established before sequencing. However, low-quality nucleic acid samples, such as Formalin-Fixed and Paraffin-Embedded (FFPE) samples, that is, FFPE samples, have many problems in building libraries. [0003] FFPE samples, as an important source of samples in the fields of biology and medicine, are widely favored by researchers. However, due to various factors, it is generally difficult to obtain high-quality and sufficient DNA from FFPE samples, especially FFPE sampl...

AI Technical Summary

Problems solved by technology

However, a large amount of DNA is lost in the purification of each step of the reaction, and the molecular diversity of the sample is affected; therefore, a high initial amount of library construction is required

However, a large number of puncture section samples or microdissected paraffin samples cannot meet the high initial volume of library construction, so it is urgent to develop an ultra-trace nucleic acid sample library construction method applied to the NGS platform

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