MDA (Multiple Displacement Amplification)-based whole-genome amplification method

Abstract

The invention provides an MDA (Multiple Displacement Amplification)-based whole-genome amplification method. The method is characterized by comprising the following steps: (1) separating and extracting single cells; (2) extracting DNA of the single cells; (3) carrying out multiple displacement amplification (MDA) by virtue of a Haplox-method single-cell whole-genome amplification kit by taking the DNA of the single cells obtained by the step (2) as a template; and (4) purifying a product obtained by the step (3) to obtain purified DNA.

Description

technical field [0001] The present invention relates to the technical field of DNA amplification, in particular to an MDA-based whole genome amplification method. Background technique [0002] In the past two decades, with the improvement of gene sequencing technology and the successive development of major international cooperation projects such as the Thousand Genomes Project, Cancer Genome Project, and Meta-Hit Project, genome research has gradually been pushed to a climax. However, the sequencing materials used so far have invariably been mixed DNA samples of millions or more cells. This method can obtain the whole genome sequence information, but the result obtained by studying it is only the average value of the signal in a group of cells, or only represents the information of the dominant number of cells, and the unique characteristics of a single cell are ignored. For example, it is nearly impossible for scientists to figure out which mutations are present in which ...

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Problems solved by technology

[0005] Among these two types, PCR amplification is more classic, but for different sequences, there is considerable deviation in the efficiency of PCR amplification, which is prone to amplification bias problems: such as CG-rich DNA sequences and corresponding genes Nonrandom loss of loci, nonrandom loss of alleles, and biases related to DNA fragment size (tend to amplify more short fragments), especially when applied to single cells where large numbers of short fragments lead to sequence loss between fragments , so that its application is limited

However, when the sample size is reduced to a single cell, accurate amplification cannot be obtained. For the single-cell genome amplification requirements, there are currently technical barriers

The MALBAC method has defects such as complex operation, low coverage, and errors introduced by PCR.

For clinical applications, there are some problems including complex process, low coverage, errors, etc.

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