Method for building high throughput sequencing library of low-quality sample deoxyribonucleic acid (DNA)

Abstract

The invention discloses a method for building a high throughput sequencing library of a low-quality sample deoxyribonucleic acid (DNA). The building method comprises the following steps: carrying out electrophoretic separation on the low-quality sample DNA, and directly obtaining the sample DNA free of a purification step; and directly building a fragmentation library from the sample DNA which is directly obtained without the purification step, so as to obtain the high throughput sequencing library. According to the building method disclosed by the invention, the fragmentation library is directly built by the sample DNA after electrophoretic separation, so that not only is the target of purifying the sample DNA achieved, but also the loss of the sample DNA is reduced, the yield of the sample DNA is improved, the quantity of the low-quality sample DNAs can meet the requirements of building the high throughput sequencing library, and pollution to protein, bacterial genome DNA or other impurities DNAs can be effectively avoided by the built high throughput sequencing library, so as to avoid the effect on following sequence assembly.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for constructing a high-throughput sequencing library of low-quality sample DNA. Background technique [0002] With the development of high-throughput sequencing technology, more and more research in the field of life sciences involves sequencing library construction and high-throughput sequencing. Sequencing library is a recombinant DNA clone population containing random fragments of all genomic DNA of an organism. It is the basis for whole genome sequencing, construction of physical map, chromosome walking, gene screening and gene map cloning, and provides a basis for genomics research. A strong technical platform. [0003] The quality of the sequencing library determines the role of the sequencing library, and the quality of the sequencing library directly depends on the quality of the sample DNA, that is, the high-throughput sequencing library has certain requirements f...

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Problems solved by technology

[0004] However, for some sample DNA mixed with other DNA impurities, sample DNA containing protein impurities, or sample DNA that is easily contaminated by host genomic DNA, such as Bacterial Artificial Chromosome (BAC) DNA, it is often difficult to purify or purify The steps are too complicated, so that the amount of purified sample DNA is less than 1ug, and the construction of high-throughput sequencing library cannot be completed, especially the small and precious sample DNA

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