Research On Detection Of Genome Variation Based On Mixed Sequencing

Genome variation is widespread in human genome,which leads to different traits among people and causes many diseases.In genomic variation,Rare Variant is a kind of single nucleotide variations with minor allele frequency less than 0.5%.Structural variation is a variation of large DNA fragments over 50 bp.With the development of sequencing technology,genome sequencing has become the mainstream method to detect rare variants and structural variation.However,due to the low allele frequency of rare variants,the detection of rare variants needs to sequence large-scale samples.However,each sample in traditional sequencing method needs to prepare DNA libraries independently,wich will cost much in DNA library preparation.In order to reduce the cost of large sample sequencing,we could use the overlapping pool sequencing.Pool sequencing can also solve the problem of structural variation detection.Traditional structural variation detection methods detect structural variations by acquiring long sequencing fragments,but overlapping pool clone sequencing can detect structural variation by acquiring haplotype information of individuals and comparing it with reference genome,which provides an accurate and economical method for structural variation detection.This paper focused on the design and test of overlapping pool sequencing on detecting rare variants and structural variations.Firstly,this paper improved the overlapping pool sequencing to detect rare variants.Based on the existing overlapping pool sequencing model,we established a more accurate decoding method was to identify the carriers of rare vairants,and evaluated the performance of overlapping pool sequencing to detect rare variants on 1000 human genome datasets.The advantages and disadvantages between traditional sequencing method and overlapping pool sequencing method were compared.The results show that the overlapping pool sequencing method can reduce the cost of sequencing to less than 50% of the common sequencing method while guaranteeing accuracy more than 99%.It has obvious advantages in detecting rare variants in large sample small fragment target sequencing.Secondly,based on the existing structural variation detection method and overlapping pool clone sequencing haplotype construction method,the information of individuals’ haplotype can be obtained,and the structural variation can be found by comparing the sequence with the reference genome.A method of genome structural variation detection for overlapping pool clone sequencing is established and the accuracy of the model is verified on the silmulated dataset.The results show that this method can reduce the cost of sequencing to less than 10% of the cost of common sequencing,on the other hand,it can detect 100% structural variation in simulated data sets,which provides a new method for structural variation detection.This paper improved and designed a new method of overlapping pool sequencing to detect rare variants and structural variations,and verified the accuracy of the method through related experiments.It expanded the application of overlapping pool sequencing and provided a new strategy to detect rare and structural variations.