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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 unfavorable RNA solution absorption, loss of RNA activity, RNA loss, etc., and achieve the effect of low extraction cost, good repeatability, and simple operation steps

Inactive Publication Date: 2006-11-15
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 treatment)

[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 37C for 12 hours, and sterilized at 125℃ 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 continuou...

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Abstract

The invention discloses a method for rapidly extracting total RNA from lotus root tissue. The steps are: first, the preparation of experimental medicines and the treatment of experimental supplies; second, the collection of materials, the young leaves of lotus root and the petioles connected with the leaves; the third, extraction, precipitation, and centrifugation to obtain total RNA; the fourth is The identification of total RNA was detected by UV spectrophotometer and formaldehyde-denatured agarose gel electrophoresis respectively. According to the obtained data and electrophoresis results, it was shown that the obtained lotus root tissue total RNA had good integrity, high purity and high yield. The invention has the advantages of simple and convenient experimental process, convenient and safe operation, accurate and effective results and good repeatability, and the method is also suitable for extracting total RNA from plant tissues of other species rich in polysaccharides and polyphenols.

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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Patent Type & Authority Patents(China)
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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