A pre-application mutation detection method for induced pluripotent stem cells

Abstract

The invention relates to an induced totipotential stem cell early-stage application mutation detection method. The method comprises the steps of selecting multiple samples and one reference gene, acquiring the sequencing data of the multiple samples, comparing the sequencing data of the multiple samples with the reference gene, and generating bam files of the multiple files correspondingly; removing deviation caused by PCR amplification in the bam files of the multiple samples, and generating corrected bam files of the multiple samples; acquiring pileup files in the corrected bam files of the multiple samples; conducting pairwise comparison on the pileup files of the multiple samples, and acquiring multiple groups of comprehensive variation detection information between every two different samples; filtering the multiple groups of variation difference information between every two different samples based on the predetermined filtration condition to obtain a result containing heterozygosis and homozygosis variation information of multiple groups of samples. Through comparison of original data, data filtration is conducted according to sequencing reads cover depth, and mutation site information can be detected more comprehensively.

Description

technical field [0001] The invention relates to a method for analyzing biological information, in particular to a method for detecting mutations in the early stage of induced pluripotent stem cell application. Background technique [0002] The main software currently used for variant screening in high-throughput sequencing results includes samtools and GATK. GATK is mainly used for mutation screening in sequencing data, including single nucleotide polymorphism (SNP) and deletion insertion (Indel), etc., and data analysis is generally performed through the process of BWA+GATK. Samtools is a collection of tools for manipulating sam and bam files, including many commands, one of which is mpileup command for generating bcf files, and then using bcftools for SNP and Indel analysis. However, whether it is the bcftools of samtools or the GATK analysis process, in order to find high-quality variation information, the analysis principle involves a series of data filtering processes....

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Problems solved by technology

This makes it impossible to obtain all the mutation information through the above method

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