High-flux low-cost molecular identification technique of trace biological samples

Abstract

The invention relates to the field of molecular organisms, in particular to a high-flux low-cost molecular identification technique of trace biological samples. The high-flux low-cost molecular identification technique of trace biological samples comprises contents in two respects of high-quality genome extraction and sequence detection and analysis. The invention aims to reduce high-flux sequencing cost of molecular identification of the trace biological samples. The high-flux low-cost molecular identification technique of trace biological samples comprises the following steps of (1) collecting samples; (2) splitting the collected samples; (3) collecting nucleic acid; (4) performing PCR amplification on marked primers; and (5) performing mixed sequencing and position reduction on PCR products. A complicated protoplast does not need to be prepared, receptor sources are simple, and the high-flux low-cost molecular identification technique of trace biological samples is suitable for transformation of fungi without spores. In addition, an existing DNA extraction technique is simplified and improved, a researcher can utilize some simple small-scale equipment in a laboratory, and large-scale high-quality genome extraction is convenient to develop; and besides, ordinary PCR and a high-flux sequencing technique are integrated, computer analysis is combined, and low-cost PCR sequence amplification and sequencing analysis of extracted DNA can be realized.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and relates to a high-throughput and low-cost molecular identification technology of trace biological samples, including high-quality genome extraction and sequence detection and analysis of trace biological samples. Background technique [0002] Modern molecular biotechnology has gradually entered the era of high-throughput processing in DNA sample extraction, PCR amplification and sequencing, which is very necessary in many research fields, such as studying the diversity of certain biological loci by molecular means When processing a large number of samples (including animal and plant cells and microorganisms), such high-throughput processing usually relies on expensive automated processing machines and kits. For laboratory research with ordinary conditions, if the genome extraction, PCR amplification and sequencing are performed manually one by one with kits according to conventional...

AI Technical Summary

Problems solved by technology

In recent years, many researchers have proposed many improvements in DNA extraction methods, but they are often only aimed at one type of microbial sample, such as only for Gram-negative bacteria, which makes it difficult to extract various samples at the same time. For example, intestinal bacteria include Gram-negative bacteria, which are relatively easy to extract DNA, and Gram-positive bacteria, such as Bifidobacteria, which are difficult to extract DNA; moreover, in order to meet the needs of large-scale extraction, the previous large-scale Most improvements tend to oversimplify the extraction process, seriously reducing the quality of the extracted DNA

In terms of sequence detection and analysis, although PCR and classical sequencing methods are very mature and reliable, it is still time-consuming and labor-intensive to sequence thousands of samples, making high-throughput sequencing an astonishing amount of sequencing, but the cost of sequencing each sample Not cheap, expensive to use