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Quick extracting method for lotus rhizome tissue total RNA

An extraction method and tissue technology, applied in the field of rapid extraction of total RNA from lotus root tissue, can solve the problems of RNA activity loss, RNA loss, unfavorable RNA solution absorption, etc., and achieve the effect of low extraction cost, good repeatability, and wide applicability

Inactive Publication Date: 2005-03-02
WUHAN UNIV
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Problems solved by technology

Polysaccharides form insoluble jelly and co-precipitate with RNA; while phenolic compounds are easily oxidized to brown substances, and then irreversibly combine with RNA, resulting in loss of RNA activity and loss of RNA when extracted with chloroform, or Form insoluble complexes; and the floating fat layer during the separation process is also not conducive to the absorption of RNA solution

Method used

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  • Quick extracting method for lotus rhizome tissue total RNA

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Embodiment Construction

[0026] 1. Preparation of experimental drugs and handling of experimental supplies:

[0027] (1), the preparation of experimental drugs:

[0028] RNA extraction buffer: 2% CTAB (w / v), 2-5% PVP K30 (w / v), 100mM Tris-Cl (PH8.0), 25mM EDTA (PH8.0), 2M NaCl, 2-5% β-ME (add when used)

[0029] 10M LiCl (with 0.1% DEPC-H 2 O treatment)

[0030] 3M NaAc(PH5.2) (with 0.1% DEPC-H 2 O processing)

[0031] 96% ethanol (RNase-free H 2 O preparation)

[0032] 75% ethanol (RNase-free H 2 O preparation)

[0033] "Chloroform:isoamyl alcohol (24:1)"

[0034] (2) Handling of experimental supplies:

[0035] All the solutions in the RNA extraction except containing Tris, all use 0.1% DEPC-H 2 O prepared, incubated at 37°C for 12 hours, and sterilized at 125°C for 30 minutes; the solution containing Tris was treated with DEPC-treated RNase-free H 2 O, prepared directly after autoclaving. Glass, pottery, metal and other utensils that can withstand high temperature should be baked contin...

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Abstract

The quick extracting process of lotus rhizome tissue total RNA includes the steps of: compounding test chemical preparation and processing test articles; sampling tender leaf and leaf stalk; extracting, precipitating and centrifuging to obtain total RNA; identifying total RNA with ultraviolet spectrophotometer and formaldehyde denatured agarose gel electrophoresis detection. The obtained data and electrophoresis results show that the lotus rhizome tissue with complete total RNA has high purity and high yield. The present invention has simple test process, convenient and safe operation, accurate and effective result and high repeatability. The method is also suitable for the total RNA extraction of other plant tissue with rich polysaccharide and polyphenol.

Description

Technical field: [0001] The present invention relates to the molecular biology experiments of lotus root gene cloning, gene function and transgenic lotus root, and more specifically relates to a rapid extraction method of total RNA from lotus root tissue, which is suitable for the extraction of total RNA from lotus root and other plant tissues rich in polysaccharides and phenolic compounds . Background technique: [0002] Lotus root is an important aquatic economic crop. It is not only edible, but also an important ornamental flower and Chinese herbal medicine. The research on it has attracted widespread attention. As far as the genetics of lotus root is concerned, the traditional research is mainly from the morphological aspect, and the recent research is mainly on the molecular aspect by using the method of molecular biology. [0003] Plant total RNA extraction technology is a basic experimental technique of plant molecular biology and a necessary prerequisite for plant m...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12N15/29
Inventor 王曼玲朱虹玲周立胡中立周明全宋运淳
Owner WUHAN UNIV
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