Screening method of molecular reverse probes for capturing specific region of enriched genome

Abstract

The invention discloses a screening method of molecular reverse probes for capturing a specific region of an enriched genome. The method comprises the steps that firstly the structures of the molecular reverse probes are designed; starting from the 5' end of the target region, the probes with at least 50% of scanned areas falling on the 5' end are selected to serve as a candidate starting set; adjacent probes are scored in pairs, the probes meeting the requirement are combined with the previous selected probes sequentially to form a set of all possible probe combinations, each candidate combination is scored, score calculation is conducted on all possible combinations, and the combination with highest scores or lowest penalty scores is obtained. The method is applicable to linker structures of different sequencing platforms, an obtained molecular reverse probe technique is remarkably improved at the aspects of precision, convenience, detection sensitivity and the like, meanwhile, a coverage target region, especially the number of the probes needed by a klenow fragment exon region is effectively reduced, and the method is convenient and rapid, and saves the cost.

Description

technical field [0001] The invention relates to the field of genetic engineering probes, in particular to a screening method for molecular reverse probes used to capture and enrich specific regions of genomes. Background technique [0002] Rare mutations and de novo mutations are the genetic basis of complex diseases. It is of great significance to detect the genetic diversity in a sample cell population for the study of human cancer genetics. However, despite the decreasing cost and increasing throughput of massively parallel sequencing, it is still very challenging to stably detect mutations with low relative abundance in a specific DNA sample. [0003] Many methods have been used to identify gene mutations, including qPCR, mass spectrometry, microarray chips, etc. However, these methods are not very sensitive to the identification of low-frequency mutations. Therefore, there is an urgent need for a method that can handle a large number of DNA molecules and can more sensi...

AI Technical Summary

Problems solved by technology

The traditional design method mainly considers the specificity of the primers at both ends to select, and less consideration is given to the optional range and experimental characteristics of the probes

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