Method for evaluating STR slippage by molecular specific bar code in next-generation sequencing platform and application of method
A next-generation sequencing, specific technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., to reduce the effect of STR slippage
Pending Publication Date: 2022-01-14
JIANGHAN UNIVERSITY
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[0007] This application provides a method for assessing STR slippage using molecular-specific barcodes in a next-generation sequencing platform and its application to solve the technical problem of how to slip STR in a next-generation sequencing platform
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[0048] In order to make the purposes, technical solutions and advantages of the embodiments of the present application more clear, the technical solutions in the embodiments of the present application will be clearly and completely described below in conjunction with the drawings in the embodiments of the present application. Obviously, the described The embodiments are some of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present application.
[0049] The excellence of the method of the present application will be specifically described below in conjunction with the examples.
[0050] The method for evaluating the slippage of STR in next-generation sequencing using molecular-specific barcodes is characterized in that the method comprises the following steps: ...
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The invention relates to the field of next-generation sequencing, in particular to a method for evaluating STR slippage by using a molecular specific bar code in next-generation sequencing and application of the method. The method comprises the steps of enabling the two ends of a DNA fragment, after the genome of a sample to be detected is broken, to be connected with excessive molecular specific bar code sequences correspondingly, and performing PCR amplification to obtain an original DNA template molecule containing a sample specific identification code and UMI; performing STR target sequence capture to obtain a captured target fragment; performing high-throughput sequencing on the captured target fragment to obtain sequencing data; performing STR typing on the sequencing data according to a UMI linker sequence, a sample specific identification code and a reference genome sequence of the to-be-detected sample to obtain the slippage rate of each STR gene locus; and evaluating the slippage of the STR in a next-generation sequencing platform. The UMI can accurately recognize and remove the STR locus stutters in the PCR process, the expected effect of reducing STR slippage in a next-generation sequencing platform is achieved, and technical support is provided for fine identification of mixed samples in forensic identification.
Description
technical field [0001] The present application relates to the field of next-generation sequencing, in particular to a method for evaluating STR slippage by using molecular-specific barcodes in next-generation sequencing and its application. Background technique [0002] Short tandem repeats (short tandem repeats, STRs) are 2 to 6 nucleotides as repeating units, which are arranged in tandem repeats in the human genome. The length of STR gene loci is generally between 100 and 300 bp, and the copy number and sequence of STR core sequence among individuals are highly polymorphic. [0003] At present, STR typing has been widely used in the fields of forensic individual identification and paternity testing. [0004] The classic principle of STR typing is to design site-specific specific primers using the conserved sequences in the flanking regions of STRs, amplify them on a PCR instrument, and finally detect the amplified products by electrophoresis. The STR typing technology ba...
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IPC IPC(8): C12Q1/6869C12Q1/6806
CPCC12Q1/6869C12Q1/6806C12Q2535/122C12Q2537/165C12Q2525/151C12Q2565/519C12Q2525/191Y02A90/10
Inventor 彭海陈利红李论李甜甜周俊飞高利芬方治伟肖华锋
Owner JIANGHAN UNIVERSITY