Efficient extraction method of guava fruit RNA

Abstract

The invention belongs to the technical field of biological genetic engineering, and discloses an efficient extraction method of guava fruit RNA (Ribonucleic Acid), which comprises the following steps: preparing buffer A; preparing a total RNA extracting solution; splitting the tissues; precipitating protein; carrying out RNA precipitation; and RNA purification. Different systems can be selected according to the model of a laboratory centrifuge and the size of fruits, and the effect is stable; according to the method, polysaccharide, polyphenol and pigment which are rich in fruits can be ignored, and high-quality total RNA can be extracted. The sample amount can be extracted according to experimental materials, an extraction system can be changed, the sample amount range is wide, the method is particularly suitable for large-volume fruits, large-sample-amount extraction is selected, the representativeness is good, and the subsequent experiment repeatability is good. The concentration of the total RNA extracted from the fruits can reach 5000ng / ul, the template system added in the subsequent reverse transcription is controllable, and the reverse transcription effect is not influenced; the total RNA amount of extracted fruits is large, and the method is suitable for large-scale fluorescent quantitative screening of candidate genes.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, in particular to a method for efficiently extracting guava fruit RNA. Background technique [0002] At present, among the extraction methods for guava fruit RNA, it is difficult to extract the total RNA of fruits rich in polysaccharides and polyphenols by ordinary methods; the amount of samples extracted by ordinary methods is basically within 0.1g, which is not suitable for large-volume fruits, and 0.1g samples The representativeness is poor, and the repeatability of the experiment is poor; the concentration of total RNA extracted by common methods is low, and the concentration of extracted RNA is generally lower than 100ng / ul, and the template system added to subsequent reverse transcription is relatively large, which affects the effect of reverse transcription; The amount of RNA is small (≤5ug), which is not suitable for large-scale fluorescent quantitative screening of candidat...

AI Technical Summary

Problems solved by technology

[0004] (1) The total RNA of fruits rich in polysaccharides and polyphenols that is difficult to extract by ordinary methods, the amount of extracted samples is basically within 0.1g, which is not suitable for large-volume fruits, and the representativeness of 0.1g samples is poor, and the experimental repeatability

[0005] (2) The concentration of total RNA extracted by ordinary methods is low, and the concentration of extracted RNA is generally lower than 100ng / ul. The template system added to the subsequent reverse transcription is relatively large, which affects the effect of reverse transcription

[0006] (3) The amount of total fruit RNA extracted by ordinary methods is small (can be ≤5ug), which is not suitable for a large number of fluorescent quantitative screening candidate genes, and multiple extractions are required, with high cost and poor repeatability

[0007] The difficulty of solving the above problems and defects is: more difficult, the kit generally uses the column method, the amount of adsorbed RNA is limited, and the extraction system is small; polysaccharides and polyphenols have a greater impact on the general extraction solution, and the RNA yield is greatly affected

Even if the RNA is eluted repeatedly through the column through multi-tube extraction, the extraction amount will not increase significantly and the cost will increase

[0008] The significance of solving the above problems and defects is: to solve the problems of poor representativeness of samples, poor repeatability of subsequent quantification, etc., and obtain a large amount of high-quality RNA

Images