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A kit for detecting the expression level of soybean circadian clock gene elf3 homologous gene

A technology of gene expression and biological clock genes, which is applied in the field of expression kits, can solve the problem of high specificity and specificity of target gene primers

Active Publication Date: 2021-07-09
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it requires high specificity and specificity in the design of target gene primers, which becomes a limiting factor for accurate analysis of target gene expression levels

Method used

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  • A kit for detecting the expression level of soybean circadian clock gene elf3 homologous gene
  • A kit for detecting the expression level of soybean circadian clock gene elf3 homologous gene
  • A kit for detecting the expression level of soybean circadian clock gene elf3 homologous gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] Example 1, Preparation of a kit for detecting the expression level of homologous genes of the soybean circadian clock gene ELF3

[0081] According to the genome sequence of soybean variety Williams82, the homologous genes of soybean circadian clock gene ELF3 were obtained, including Glyma. ) and Glyma.17G231600 gene (Genebank No. 100788038).

[0082] 1. Acquisition and identification of primer pairs for detecting soybean Glyma.04G050200 gene expression

[0083] 1. Perform homologous comparison of the main transcription sequences of the Glyma.04G050200 gene, Glyma.08G197500 gene, Glyma.14G091900 gene and Glyma.17G231600 gene, select the Glyma.04G050200 gene and the other three genes with large differences, design and A primer pair was synthesized for detecting the expression level of soybean Glyma.04G050200 gene. One of the primer pairs is primer pair 200. The nucleotide sequences of the upstream primers and downstream primers constituting the primer pair 200 are show...

Embodiment 2

[0143] Example 2, using the kit prepared in Example 1 to detect the expression level of soybean ELF3 homologous gene

[0144] 1. Plant the seeds of Zhonghuang 24 in nutrient soil and cultivate them conventionally to obtain Zhonghuang 24 seedlings with fully expanded single leaves.

[0145] 2. After completing step 1, take the Zhonghuang 24 seedlings, remove the single leaves every 3 hours, freeze them thoroughly in liquid nitrogen, store them at -80°C, and use them as samples to be tested. A total of 10 days were picked.

[0146] 3. After step 2 is completed, the sample to be tested is taken, RNA is extracted with a TRNzol kit, and the RNA concentration is measured with a Nanodrop instrument to obtain the RNA of the sample to be tested.

[0147] 4. After completing step 3, take the RNA of the sample to be tested, use the TransScript One-Step gDNARemoval and cDNA Synthesis SuperMix kit for reverse transcription, and then dilute with double distilled water to obtain the sample ...

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Abstract

The invention discloses a kit for detecting the expression level of the homologous gene of the soybean biological clock gene ELF3. It includes the primer pair 200 for amplifying the Glyma.04G050200 gene, the primer pair 500 for amplifying the Glyma.08G197500 gene, the primer pair 900 for amplifying the Glyma.14G091900 gene and the primer pair for amplifying the Glyma.17G231600 gene The upstream and downstream primers of primer pair 600; primer pair 200, primer pair 500, primer pair 900 and primer pair 600 are shown in SEQ ID NO: 3 to SEQ ID NO: 10 in sequence. Experiments have shown that real-time quantitative PCR using primer pair 200, primer pair 500, primer pair 900, and primer pair 600 can obtain the expression level of the corresponding gene, and then analyze the biological rhythm and growth habit of soybean. The invention has important application value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit for detecting the expression level of a soybean circadian clock gene ELF3 homologous gene. Background technique [0002] Real-time quantitative PCR technology can conveniently and quickly evaluate the expression level of target genes, and is an important analytical tool widely used in the fields of molecular biology and molecular genetic breeding. However, it requires high specificity and specificity in the design of target gene primers, which becomes a limiting factor for accurate analysis of target gene expression levels. Especially in the case of a large number of copies of homologous genes in the genome, it is very important to obtain specific primers that can effectively distinguish each homologous gene. [0003] Soybean is a typical short-day crop and a model material for the study of plant photoperiod response. It is also the most important oil crop and high...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/166
Inventor 岳岩磊刘薇李涛王潇然郭红祥马文娅贾然然姜泽吴红霞
Owner HENAN AGRICULTURAL UNIVERSITY
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