Database-building method for amplicon sequencing

Abstract

The invention relates to the technical field of high-flux sequencing and discloses a database-building method for amplicon sequencing. The database-building method comprises the following steps of 1, object region enrichment based on PCR amplification: designing primer sequences aiming at an object region of genome DNA of a sample to be detected, wherein ends 5' of a forward primer and a reverse primer in the primer sequences are provided with random base sequences and linking sequences, carrying out PCR amplification and recovering the PCR product, and 2, sequencing linker introduction based in PCR amplification: carrying out mixing on the PCR product obtained by the step 1, designing primer sequences comprising sequences complementary with the linking sequences obtained by the step 1, carrying out PCR amplification on the mixture, and recovering the PCR product so that a DNA database for amplicon sequencing is obtained. The database-building method reduces building time and cost of the DNA database for amplicon sequencing, reduces error of bioinformatics analysis of the sequencing result and improves accuracy and authenticity of the analysis result.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a rapid library building method for amplicon sequencing. Background technique [0002] With the development of high-throughput sequencing technology and the reduction of sequencing costs, sequencing technology has gradually become a routine experimental method for basic biological research and medical testing. Since the Human Genome Project constructed the first genome map, next-generation sequencing technology has constructed genome maps of a large number of conventional species through whole genome sequencing, which has also promoted the rapid development of sequencing technology. However, the complex structure of whole genome sequencing, the large amount of data, the long cycle and the high cost limit its development. [0003] At the same time, some researchers are only interested in specific genomic regions, and in this case, they only need to perform sequencing re...

AI Technical Summary

Problems solved by technology

[0004] The current amplicon sequencing is based on conventional library construction methods. After PCR amplification and enrichment to the target area, the sample is mixed, then the end is repaired and A is added, and the adapter is added, the sample is purified, and the purpose of gel recovery is Bands, then PCR amplified, and finally run the gel to recover the main band. This conventional library construction method is cumbersome, time-consuming, and heavy workload, which is not conducive to the sequencing of a large number of samples.

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