Method for extracting total RNA from loropetalum chinense var. rubrum leaves

A technology of safflower wood and an extraction method, applied in the biological field, can solve the problems of lack of RNA extraction reagents, weak molecular mechanism research and application, limited application of modern molecular technology, etc., to achieve the effects of removing pollution and reducing RNA degradation.

Inactive Publication Date: 2013-02-06
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, there are many commercial RNA extraction reagents that can be suitable for almost all animal tissues after a small amount of adjustment, but there is no commercial RNA extraction reagent that can be suitable for most plant tissues
In addition, there is no report on the extraction of total RNA from the leaves of P. safflower, and the research and application of the molecular mechanism of leaf coloration of P. safflower are also very weak, which limits the application of modern molecular techniques in the genetic improvement of leaf color and germplasm innovation.

Method used

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  • Method for extracting total RNA from loropetalum chinense var. rubrum leaves
  • Method for extracting total RNA from loropetalum chinense var. rubrum leaves
  • Method for extracting total RNA from loropetalum chinense var. rubrum leaves

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Experimental program
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Embodiment 1

[0027] (1) Select a 10-15cm long branch with leaves from a healthy Safflower plant and quickly insert it into a beaker filled with 1 / 3-1 / 4 highly distilled water, and bring it back to the laboratory immediately;

[0028] (2) Weigh 80 mg of the leaves described in step (1), cut them into small pieces with a sterilized blade, put them into a sterilized 1.5ml centrifuge tube, soak the leaves in the centrifuge tube with liquid nitrogen for 10 seconds, and grind the leaves quickly with a small pestle into powder;

[0029] (3) Add 0.5ml plant total RNA extractant to the centrifuge tube with powdered leaves, lay the centrifuge tube flat, let it stand at room temperature for 5 minutes, centrifuge at 4°C and 12000rpm for 1 minute, and transfer the supernatant to an RNase-free centrifuge tube Wherein, the RNA extractant is purchased from RNAplant plus Reagent of Tiangen Biochemical Technology Co., Ltd., product number DP437, and β-mercaptoethanol is added before use, and the volume rati...

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Abstract

The invention discloses a method for extracting total RNA from loropetalum chinense var. rubrum leaves. The RNA extracted from the loropetalum chinense var. rubrum by adoption the method has high purity and contains a few impurities. The specific primer sequences of several commonly used internal reference genes including alpha-tubulin, beta-actin, 18S rRNA and GAPDH are used for RT-PCR (reverse transcription-polymerase chain reaction) analysis, thus a bright specific band (Figure 2) which has the same size as the designed expected fragment of the primer can be obtained. The total RNA obtained by adopting the method in the test has good integrity and can be used for the follow-up RT-PCR amplification to express and analyze related genes.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for extracting RNA from leaves of Loropetalum safflower. Background technique [0002] Loropetalum chinense var.rubrum is one of the main colorful-leaved plants with rich garden uses in the middle and lower reaches of the Yangtze River and the south of the Yangtze River. The molecular mechanism of leaf coloration and the genetic improvement of leaf color have always been of great concern to relevant researchers. problem, but isolating high-quality total RNA from Roropetus safflower is the key. During the growth process of Roropetalum safflower leaves, a large amount of tannins, reducing sugars, glycosides, flavonoids, phenolic substances and organic acids are synthesized and accumulated. These compounds will directly affect the effective extraction of RNA. [0003] In the existing plant RNA extraction, the traditional Trizol method is mainly used. However, the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 于晓英张力符红艳许威陈己任李达陈海霞
Owner HUNAN AGRICULTURAL UNIV
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