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Method for high-throughput screening of maize reference gene

An internal reference gene and high-throughput technology, applied in the field of molecular genetics, can solve the problems of insufficient screening of internal reference genes, small amount of data, and low coverage of screening internal reference genes, so as to reduce experimental workload, improve efficiency, and reduce experimental costs. cost effect

Inactive Publication Date: 2014-11-05
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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Problems solved by technology

It should be noted that the coverage of internal reference genes screened by this method is very low. It is based on the previously defined part of the housekeeping genes, and only compares the expression stability of some housekeeping genes. The amount of data is too small to screen out truly stable internal references. Gene

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  • Method for high-throughput screening of maize reference gene

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[0019] The experimental methods in the following examples are conventional experimental methods unless otherwise specified. The experimental reagents and consumables described in the following examples are from General Biochemical Reagent Company unless otherwise specified.

[0020] The method for high-throughput screening of corn internal reference genes of the present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0021] This example is a method for high-throughput screening of internal reference genes in maize. The high-throughput transcriptome sequencing (RNA- seq) database, the screening steps are as figure 1 shown, including:

[0022] (1) Use SolexaQA software to filter the RNA-seq data, remove the relatively poor quality data with a Phred value less than 20 and a length less than 20bp, and leave sequences with higher quality;

[0023] (2) Use Bowtie software to anchor the sequence obtained...

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Abstract

The invention discloses a method for high-throughput screening of a maize reference gene. The method comprises the following steps: by using a high-throughput transcriptome sequencing (RNA-seq) database which is sourced from different maize inbred lines with high diversity, different tissues and different developmental periods and is constructed after different stress treatment, analyzing the expression conditions of a whole transcriptome massively; screening a gene of which the expression is the most constant in the different developmental periods and under different conditions as the reference gene, and providing an accurate comparison for the study of transcriptome expression. The method disclosed by the invention is used for analyzing all the expressed genes of the whole transcriptome by using data of the whole transcriptome, is used for systematically screening the maize reference gene, and is more accurate and reliable compared with a conventional screening method.

Description

technical field [0001] The invention relates to the field of molecular genetics, in particular to a method for high-throughput screening of internal reference genes in maize. Background technique [0002] Gene expression refers to the conversion of genetic information stored in deoxyribonucleic acid (DNA) sequences into biologically active protein molecules during the life process of cells through transcription and translation. This process determines cell differentiation and morphological development. The amount of messenger ribonucleic acid (mRNA) produced by the transcription of each gene is regulated by various factors such as time and space, and ultimately affects the growth and development, morphological structure and biological functions of an individual. Gene regulation is one of the central topics of modern molecular biology research. In order to study the mechanism of gene expression regulation, it is necessary to understand the expression levels of different gen...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6895C12Q2600/158
Inventor 林峰赵涵
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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