Searching method for species-specific endogenous barcodes and application thereof in multi-sample mixed sequencing

Abstract

The invention discloses a searching method for species-specific endogenous barcodes and an application thereof in multi-sample mixed sequencing. The searching method comprises the following steps: determining, collecting and comparing candidate genomic sequences, calculating the variation degree of a sequence in a current sliding window and the conservation degrees of sequences on both sides of the window, and according to the scanning calculation results of the sliding window, determining an endogenous barcode; and after the endogenous barcode is determined, carrying out amplification by using an overlap extension PCR technology, connecting the endogenous barcode and a to-be-sequenced target sequence, carrying out sequencing on a computer, and determining the sample source of the sequencing segment through the characteristics of the endogenous barcode. Compared with exogenous barcode sample marked samples synthesized in vitro, endogenous barcodes do not need to be subjected to artificial synthesis of DNA, and multiple samples are simultaneously amplified in one-step reaction and connected with respective barcode and to-be-sequenced target sequence can be realized, so that an experiment process of extracting the target sequence firstly and then connecting barcodes synthesized in vitro one by one is simplified, thereby reducing the cost of sequencing.

Description

technical field [0001] The invention belongs to the field of gene sequencing, in particular to a search method for species-specific endogenous barcodes and its application in multi-sample mixed sequencing. Background technique [0002] In recent years, high-throughput sequencing technology has developed rapidly and its application scope has been continuously expanded. It is often necessary to sequence specific DNA regions in a large number of samples. In order to improve the parallel sequencing capability of multiple samples, high-throughput sequencing platforms basically provide physically segmented sequencing channels, but the parallel processing capability is limited by the number of channels, which still cannot meet the parallel sequencing requirements of a large number of samples. Therefore, multi-sample mixed sequencing The experimental plan came into being. The current main methods of hybrid sequencing include DNA barcode labeling and overlapping hybrid sequencing. ...

AI Technical Summary

Problems solved by technology

[0004] At present, in the application process of DNA barcodes, there are two operations that are time-consuming and labor-intensive: one is that DNA barcodes need to be synthesized in vitro, and the barcodes corresponding to each sample must be unique, so how many samples are mixed, how many pieces of DNA must be synthesized in vitro barcode; the second is that after the barcode is synthesized, the corresponding samples need to be connected, and this process also needs to be added one by one

When the number of samples is relatively large, the above two operations undoubtedly need to consume a lot of manpower and material resources

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