A method for high-throughput screening of differentially expressed genes in plant genomes

A high-throughput, genomics technology, applied in the field of genetics research, can solve the problems of research limitations, Sanger sequencing can not further expand parallelism and microquantification

Active Publication Date: 2021-09-24
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, Sanger sequencing has limited further research due to its inability to further expand parallelism and microquantification.

Method used

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  • A method for high-throughput screening of differentially expressed genes in plant genomes
  • A method for high-throughput screening of differentially expressed genes in plant genomes
  • A method for high-throughput screening of differentially expressed genes in plant genomes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Raw material acquisition: Populus tomentosa '1316' is an excellent clone of Populus tomentosa from the National Germplasm Bank of Populus tomentosa;

[0064] All reagents used in the CTAB method are commercially available;

[0065] The specific operation steps are as follows:

[0066]Step S101, select the annual plants of excellent clones of Populus tomentosa, and plant the clone plants used in the experiment in flower pots with soil, peat and perlite (volume ratio of 1:1:1) as the substrate, and the place is Beijing Forestry University Greenhouse (40°0'N, 116°20'E), the light time is 16 hours per day, and the temperature is 20°C. Adopt the IAA solution of 100 μ M (IAA powder is dissolved in 95% ethanol, then constant volume to 100 μ M with distilled water.) The leaves of the Populus tomentosa clone plant of the experimental sample are sprayed until the liquid drips on the leaves; the control sample is sprayed with the same Method of spraying with water. The leaves o...

Embodiment 2

[0108]The following will take the screening process of differential expression of alleles before and after drought stress treatment (taken from the National Populus microphylla germplasm resource bank) as an example to further illustrate the screening method provided by the present invention.

[0109] The acquisition of raw materials: select individuals with the same or similar growth potential in the Populus small-leaved clones, and carry out the following treatments: drought treatment, no watering for 10 days; control samples, watering according to the normal cultivation time (watering once every two days, until the soil is fully drenched).

[0110] All reagents used in the CTAB method are commercially available;

[0111] The specific operation steps are as follows:

[0112] Step S101, select excellent clone plants of Populus microphylla, and plant the clone plants used in the experiment in flower pots with soil, peat and perlite (1:1:1 volume ratio) as substrates, and the ...

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Abstract

The invention provides a method for high-throughput screening of heterozygous variant sites expressed in allelic imbalance in plant genomes, which belongs to the field of genetics research and comprises the following steps: 1) processing plant materials with exogenous regulatory factors to obtain processed samples; 2) ) extract the total RNA of the processed sample and the control sample, construct a strand-specific sequencing library and perform high-throughput sequencing; 3) filter the raw sequencing data to obtain clean reads; 4) compare the clean reads with the reference genome respectively Obtaining the alignment result; 5) deleting redundant duplicate reads in the alignment result, and re-aligning the sequences around the INDEL site to obtain the final alignment result data; 6) performing allelic positioning on the final alignment result data Point variant screening obtained heterozygous variant sites with allelic imbalance expression. The method solves the analytical challenge of resolving the response of the genome to the external environment at the allelic level.

Description

technical field [0001] The invention belongs to the field of genetics research, in particular to a method for high-throughput screening of genes differentially expressed in plant genomes. Background technique [0002] With the development of sequencing technology, high-throughput sequencing technology has become more and more widely used, providing important information for biological research such as revealing genetic information and gene expression regulation, and has become the most commonly used experimental technology for genomics research. Since 1973, the Sanger sequencing method has developed rapidly due to its advantages of convenience, simplicity, reliability and accuracy, and long sequencing fragments, and has been widely used in scientific research, making important contributions to scientific development. However, Sanger sequencing has limited further research because of its shortcomings such as inability to further expand parallelism and microquantification. Hi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6869
CPCC12Q1/6869C12Q1/6895C12Q2600/156C12Q2535/122
Inventor 张德强轩安然宋跃朋
Owner BEIJING FORESTRY UNIVERSITY
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