Medium-throughput gene expression analysis method based on second-generation test platform

Abstract

The invention relates to a medium-throughput gene expression analysis method based on a second-generation test platform. The medium-throughput gene expression analysis method comprises steps as follows: corresponding PCR (polymerase chain reaction) primers are divided into two groups according to the relative expression quantity of multiple genes, and the concentration proportion of primers is adjusted in the groups; competitive templates of standard substances and a competitive template of a to-be-tested sample are established, and three rounds of multiple competitive PCRs (polymerase chain reactions) are performed; reaction products of the three rounds of reactions are mixed to serve as a second-generation testing platform library for computer sequencing, data are extracted and analyzed, and the difference of relative expression quantities of multiple genes in different samples can be obtained. On the basis of the high-speed-developed second-generation sequencing platform, a target template and an internal reference template are accurately and rapidly quantified, expression difference analysis of five target genes can be performed on multiple samples simultaneously, the procedure is simple and easy to implement, and the cost is low; when multiple gene expression differences of multiple samples are required to be analyzed, the method can be a high-accuracy, moderate-throughput and low-cost method.

Description

technical field [0001] The invention belongs to the technical field of gene expression analysis, in particular to a medium-throughput gene expression analysis method based on a second-generation test platform. Background technique [0002] In the post-genomics era, the focus of genomics research has shifted from revealing all the genetic information of life to the study of functions at the molecular level. By analyzing gene expression levels, researchers can obtain relevant information such as gene transcription, gene regulation, signal transduction pathway, nucleic acid, protein structure, function and their interrelationships; reveal gene expression and regulation patterns in time and space, and provide a basis for studying gene function. , providing new methods for elucidating complex traits. [0003] Commonly used techniques for analyzing the expression level of target genes are as follows: traditional Northern blot analysis and ribonuclease protection assay (Ribonuclea...

AI Technical Summary

Problems solved by technology

In the analysis of gene expression, next-generation sequencing platforms have also been applied, such as digital gene expression profiling (RNA-seq) based on transcriptome sequencing, but it is difficult to distinguish many low-abundance and Differential information of medium-abundance genes (SimonJ.Furney, MalinPedersen.(2013) CANCERDISCOVERY10, 1122-9), and researchers often only need to analyze the expression difference of the genes they are interested in. Therefore, a highly accurate High-speed, moderate-throughput, and low-cost gene expression analysis methods need to be established urgently

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