Tropical plant polysaccharide and polyphenol small RNA extraction method

A tropical plant and extraction method technology, applied in the field of RNA extraction, can solve problems such as mobility changes, and achieve the effects of simple operation, rapid extraction and detection analysis, and wide application prospects

Active Publication Date: 2014-04-23
ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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Problems solved by technology

[0010] The purpose of the embodiments of the present invention is to provide a method for extracting small RNA polysaccharides and polyphenols from tropical...

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  • Tropical plant polysaccharide and polyphenol small RNA extraction method
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  • Tropical plant polysaccharide and polyphenol small RNA extraction method

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[0053] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0054] The application principle of the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments.

[0055] like figure 1 As shown, the tropical plant polysaccharide polyphenol small RNA extraction method of the embodiment of the present invention comprises the following steps:

[0056] S101: Sampling leaves of tropical plants, freezing them in liquid nitrogen, and storing them at -80°C;

[0057] S102: Prepare small RNA extraction reagents, CTAB lysis buffer, PEG8000 precipitation buffer, EDC cross-linking buffer, miRNA Northern blotting hybridization buffer, ...

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Abstract

The present invention discloses a tropical plant polysaccharide and polyphenol small RNA extraction method, which comprises: sampling tropical plant leaves, freezing in liquid nitrogen, and storing at a temperature of -80 DEG C; preparing small RNA extraction reagents, wherein the reagents comprise a CTAB lysis buffer solution, a PEG8000 precipitation buffer solution, an EDC cross-linking buffer solution, a miRNA Northern blotting hybridization buffer solution, 7% dPAGE, and Washing Buffer; according to small RNA extraction pretreatment, Trizol extraction, fractional precipitation of high molecular weight RNA, and small RNA precipitation, extracting small RNA; according to 17% dPAGE separation, trarsmembrane and cross-linking, miRNA hybridization and washing of membrane, small RNA hybridization and washing of membrane, and exposure and storage, achieving RNA hybridization; carrying out miRNA stem-loop RT-PCR amplification; and carrying out extraction and detection analysis on polysaccharide and polyphenol plant small RNA. According to the present invention, the defects in the current plant small RNA extraction method are overcome, characteristics of fastness, stability and easy operation are provided, rapid extraction and detection analysis can be performed on miRNA and siRNA, and broad application prospects are provided.

Description

technical field [0001] The invention belongs to the technical field of RNA extraction, and in particular relates to a method for extracting tropical plant polysaccharide polyphenol small RNA. Background technique [0002] Plant small RNAs are negative regulators that regulate gene expression, DNA and histone methylation and resistance to exogenous genetic factors such as viruses, transposons or transgenes at the transcriptional and post-transcriptional levels according to sequence specificity. According to the biosynthesis characteristics of small RNA, plant small RNA is mainly divided into two categories: plant small interfering RNA (siRNA, small / short interfering RNA) and microRNA (miRNA, miRNA). The length of the two is about 20-25nt, both are processed by the RNAe III-like endonuclease Dicer (or Dicer-like protein), and their formation is both related and different. MicroRNA (or miRNA) is a single-stranded small molecule RNA with a length of 20-24nt, which is produced b...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 彭军张贺蒲金基张欣谢艺贤漆艳香
Owner ENVIRONMENT & PLANT PROTECTION INST CHINESE ACADEMY OF TROPICAL AGRI SCI
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