Method for screening real-time fluorescence quantification PCR internal reference molecules of syntrichia caninervis in desert

A technology of real-time fluorescence quantification and E. denticola, which is applied in biochemical equipment and methods, microbe determination/inspection, etc., can solve problems such as blank research and screening of internal reference molecules, and unreported sequence data.

Inactive Publication Date: 2014-06-18
XINJIANG INST OF ECOLOGY & GEOGRAPHY CHINESE ACAD OF SCI
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Problems solved by technology

The characteristics and functions of many anti-stress genes need to be studied urgently, but there is no report on the sequence data of the internal reference genes of this species, and the research and screening of the most suitable internal reference molecules are even blank

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  • Method for screening real-time fluorescence quantification PCR internal reference molecules of syntrichia caninervis in desert
  • Method for screening real-time fluorescence quantification PCR internal reference molecules of syntrichia caninervis in desert
  • Method for screening real-time fluorescence quantification PCR internal reference molecules of syntrichia caninervis in desert

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Embodiment Construction

[0278] In the following examples, it is carried out according to routine experimental operations, such as "Refined Molecular Biology Experiment Guide" (F.M Osper, R.E. Kingston, J.G Seidman editor-in-chief, Ma Xuejun, Shu Yuelong translation, Beijing: Science Press, 2004 ) method described in;

[0279] Selection of internal reference molecules:

[0280] Researching and screening the most stable internal reference molecules and the most suitable internal reference items (or the optimal internal reference combination) in specific plants are the prerequisites for fluorescent quantitative PCR experiments. At present, the internal references used in the literature for analysis of plant gene expression mainly include 18SrRNA, Actin, GAPDH and other classic housekeeping genes reported in previous literatures. Now more and more reports prove that traditional classic housekeeping genes are not as stable as imagined, so many attempts on new internal reference genes have been carried ou...

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Abstract

The invention relates to a method for screening real-time fluorescence quantification PCR internal reference molecules of syntrichia caninervis in a desert. The method comprises the following steps: selecting 15 internal reference candidate genes by utilizing a syntrichia caninervis transcriptome database, designing an internal reference gene specific primer of real-time fluorescence quantification PCR by taking 15 internal reference genes as templates; carrying out a fluorescence quantification PCR experiment by selecting syntrichia caninervis gametophytes stressed by 10 non-living things and free of stress (in contrast) as experimental materials; carrying out fluorescence quantification data analysis by using geNorm, NormFinder and Refinder software, so as to screen out the best fit and the most stable internal reference molecules CDPK and alpha-TUB2 of the syntrichia caninervis for developing fluorescence quantification research under various abiotic stresses. By adopting the method disclosed by the invention, the errors of the syntrichia caninervis transcriptome material under different abiotic stresses in ribonucleic acid (RNA) quality, yield, reverse transcription efficiency and the like can be avoided, the real-time fluorescence quantification detection data can be better corrected and standardized, and the accuracy and the reliability of the gene quantification research are improved.

Description

technical field [0001] The invention relates to a method for screening internal reference molecules suitable for real-time fluorescence quantitative PCR research under various abiotic stresses of the desert moss Erythrodonticus. Background technique [0002] Drought, high salinity and other abiotic stresses are the main factors limiting plant growth and crop yield, resulting in more than 50% crop yield reduction every year. With global warming, the intensification of the greenhouse effect and land desertification, the problem of drought will become increasingly prominent, and the impact of drought on agricultural production will also be more serious. At present, the use of drought stress-related genes to improve the drought resistance of plants has become a research hotspot in plant stress resistance molecular biology and an important research direction in plant stress resistance genetic engineering. [0003] The drought-resistant gene resources accumulated by plants in ari...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q2561/113C12Q2545/101
Inventor 张道远李小双李海燕杨红兰
Owner XINJIANG INST OF ECOLOGY & GEOGRAPHY CHINESE ACAD OF SCI
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