Sugarcane genome endogenous reference gene acetolactate synthase gene PCR (polymerase chain reaction) primer sequences and amplification method

A technology of acetolactate synthase and internal standard gene, applied in recombinant DNA technology, DNA/RNA fragments, DNA preparation, etc., can solve the problem of no research report on copy number, achieve easy judgment, ensure amplification efficiency, and applicability wide effect

Inactive Publication Date: 2015-07-08
FUJIAN AGRI & FORESTRY UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

There have been many studies on the acetolactate synthase gene that produces this enzyme, but there is no research report on the copy number of this gene in the genome of sugarcane species

Method used

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  • Sugarcane genome endogenous reference gene acetolactate synthase gene PCR (polymerase chain reaction) primer sequences and amplification method
  • Sugarcane genome endogenous reference gene acetolactate synthase gene PCR (polymerase chain reaction) primer sequences and amplification method
  • Sugarcane genome endogenous reference gene acetolactate synthase gene PCR (polymerase chain reaction) primer sequences and amplification method

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Experimental program
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Embodiment 1

[0021] (1) PCR amplification of the internal standard gene acetolactate synthase gene in the sugarcane genome

[0022] Acetolactate synthase gene PCR primer sequence is: ScALS-F : CTCCCAGTGAAGGTCTTTGCG; ScALS-R : TGCTGGAATGTTGAACCCTTTT.

[0023] PCR reaction system: 1.0 U Taq DNA polymerase, 1×PCR buffer solution, 0.25 mM dNTP, 3.0 mM MgCl2, primers ScALS-F with ScALS-R Each 0.25 mM, DNA template 25 ng, the total volume of the reaction system is 25??l;

[0024] PCR amplification conditions: pre-denaturation at 95°C for 6 min; denaturation at 94°C for 30 s; annealing at 65°C for 30 s, extension at 72°C for 35 s, a total of 35 cycles, and a final extension at 72°C for 6 min.

[0025] (2) Electrophoresis detection of PCR product of sugarcane acetolactate synthase gene and analysis of product specificity and generality

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Abstract

The invention discloses sugarcane genome endogenous reference gene acetolactate synthase gene PCR (polymerase chain reaction) primer sequences and an amplification method. The primer sequences are composed of ScALS-F and ScALS-R. The PCR amplification method comprises a reaction system and amplification conditions. The invention provides a genome endogenous reference gene for PCR detection and species specific identification of transgenic sugarcane, and meanwhile, provides insurance for detection precision and result accuracy of the transgenic sugarcane.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a PCR primer sequence and an amplification method of an internal standard gene acetolactate synthase gene in sugarcane genome. Background technique [0002] Transgenic technology is becoming more and more mature, and the types of genetically modified sugarcane are becoming more and more diverse. At present, herbicide-resistant, insect-resistant, and disease-resistant genetically modified sugarcane has been field tested in 14 countries including the United States, Australia, India, and Brazil, creating conditions for the commercial cultivation of genetically modified sugarcane. In my country, sugarcane gene cloning and transgenic research only started in the mid-1990s. Although it started late, it has made rapid progress. The transgenic sugarcane that has been obtained includes insect-resistant genetically modified sugarcane, drought-resistant genetically modified ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/10
Inventor 陈平华林冰陈忠伟陈利平施桂姣张卓项慰刘迪夏峰王恒波高三基许莉萍陈如凯
Owner FUJIAN AGRI & FORESTRY UNIV
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