Unicellular whole genome amplification and library construction method capable of improving coverage degree of genome

Abstract

The invention provides a unicellular whole genome amplification method capable of improving coverage degree of a genome and a library construction method. According to the unicellular genome amplification method, a unicellular genome is amplified with a multiple displacement amplification method; the method comprises steps as follows: a group of primers, DNA polymerase and a unicellular genome arecontacted in a solution; the solution is amplified, so that the unicellular genome is subjected to an amplification reaction, wherein the amplification time lasts for 0.5-2 h.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a single-cell whole-genome amplification and second-generation sequencing library construction method capable of improving genome coverage. Background technique [0002] Single-cell sequencing technology (single-cell sequencing) was awarded the annual technology of "Nature Mathods" in 2013, and has broad application prospects in the fields of cancer, assisted reproduction and immunology. [0003] Single-cell genome sequencing is a technique for amplifying and sequencing the whole genome at the single-cell level. High-throughput sequencing for revealing cell population differences and cell evolution relationships. [0004] The genome of a single cell is extremely small. In order to meet the requirements of library quality control, on-machine sequencing and other detection, whole genome amplification must be performed to achieve linear or exponential growth of the geno...

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Problems solved by technology

However, in terms of genome coverage, the above three single-cell whole-genome amplification methods are not ideal, the highest is only about 60%, and there is a trend of MDA>MALBAC>DOP-PCR [4]

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