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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, and achieve easy determination, strong specificity, and wide applicability. Effect

Inactive Publication Date: 2013-11-27
FUJIAN AGRI & FORESTRY UNIV
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AI Technical Summary

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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Embodiment 1

[0022] (1) PCR amplification of the internal standard gene acetolactate synthase gene in the sugarcane genome

[0023] Acetolactate synthase gene PCR primer sequence is: ScALS-F : CTCCCAGTGAAGGTCTTTGCG; ScALS-R : TGCTGGAATGTTGAACCCTTTT.

[0024] PCR reaction system: 1.0 U Taq DNA polymerase, 1×PCR buffer solution, 0.25 mM dNTP, 3.0 mM MgCl2, primers ScALS-F and ScALS-R Each 0.25 mM, DNA template 25 ng, the total volume of the reaction system is 25??l;

[0025] 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.

[0026] (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 present invention is a biotechnology field, which involves a serial sequence and amplification method of a navigated acetylcelase genes in the sugarcane genome. Background technique [0002] GM technology is becoming more and more mature, and the types of genetically modified sugarcane are becoming increasingly diverse.At present, the antidine, insect -resistant, and disease -resistant genetically modified sugar cane has conducted field trials in 14 countries including the United States, Australia, India, and Brazil, creating conditions for the commercialized cultivation of genetically modified sugar cane.In our country, the glucan gene cloning and genetically modified research began in the mid -1990s. Although the start is late, it has made rapid progress.Sugarcane, antiviral transgenic sugarcane, antifungal genetically modified sugarcane, antidible genetically modified sugarcane, etc.With the continuous increase of genetically modified crops worldwide...

Claims

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Application Information

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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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