High-quality high-efficiency polyphenol polysaccharide plant sample RNA (ribonucleic acid) extraction method

An extraction method and polysaccharide technology, applied in the field of molecular biology, to achieve the effect of high purity

Active Publication Date: 2014-09-03
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Aiming at the demand for high-quality plant RNA in current molecular biology research, the present invention d

Method used

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  • High-quality high-efficiency polyphenol polysaccharide plant sample RNA (ribonucleic acid) extraction method
  • High-quality high-efficiency polyphenol polysaccharide plant sample RNA (ribonucleic acid) extraction method
  • High-quality high-efficiency polyphenol polysaccharide plant sample RNA (ribonucleic acid) extraction method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Extraction of Yellow Beam Leaf RNA

[0018] Take the middle and upper mature leaves of the annual Huangliangmu (plant height 1.5m, ground diameter 2.1cm), quickly put them into liquid nitrogen for quick freezing, grind them into tiny particles and put them into 1.5ml RNase-free centrifuge tubes, each tube sample is about 100mg .

[0019] The pre-prepared CTAB lysate was preheated at 60°C, pipetted 600 μl into the sample, and added 10 μl of β-mercaptoethanol at the same time, vortexed for two minutes, then lysed at 60°C for 10 minutes at a constant temperature, and mixed several times during the period.

[0020] Add 600 ul of chloroform:isoamyl alcohol extract mixed at 24:1, vortex for 2 min, and centrifuge at 14,000 x g for 10 min at 4°C. Aspirate the supernatant into a new centrifuge tube, add 600ul of the extract again, vortex for 2min, and centrifuge at 14,000xg at 4°C for 10min.

[0021] Aspirate the supernatant into a new centrifuge tube, add 500ul of B...

Embodiment 2

[0034] The extraction of embodiment two yellow beam wood leaf bud RNA

[0035] In this example, the time of chloroform in step 1 of Example 1: isoamyl alcohol extraction and centrifugation was changed from 10 min to 20 min, and the others were the same.

[0036] The test results of the above examples are shown in the accompanying drawings.

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Abstract

The invention relates to a polysaccharide polyphenol plant sample RNA (ribonucleic acid) extraction method. By combining a CTAB (cetyltrimethylammonium bromide) process and a commercial kit provided by an OMEGA kit, the invention develops a high-efficiency high-quality RNA extraction method. The inspection proves that the method is applicable to various tissues of Neolamarckia cadamba, Camellia sinensis L., Eriobotrya japonica Lindl., Rosa chinensis, Litchi chinensis Sonn., Ginkgo biloba L., Opuntia ficus-indica L., Pinus taeda L., Aloe barbadensis Mill., Taxus media and other plants, and provides a favorable option for RNA extraction and preparation in molecular biology experiments in future, and the effect is good.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, in particular, the invention relates to a method for extracting plant RNA with high efficiency and high quality. Background technique [0002] Extracting high-quality RNA without contamination from polysaccharides, polyphenols, genomic DNA, proteins, or other secondary metabolites is crucial for many follow-up experiments such as studying the function and expression of genes during plant growth and development, such as inversion Recording PCR, real-time quantitative PCR, cDNA library construction, Northern blot hybridization and RNA-seq, etc. [0003] Phenols are easily oxidized to quinones, which irreversibly bind to proteins and nucleic acids. However, polysaccharides can co-precipitate with RNA, reducing the purity of RNA and affecting the use in downstream experiments. [0004] Commercial RNA extraction kits commonly used in the market today, such as Takara, Promega, and Omega, al...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 李俊成欧阳昆唏黄浩陈晓阳
Owner SOUTH CHINA AGRI UNIV
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