The invention provides an extration method of
plant RNA by utilizing guanidinium
isothiocyanate-
chloroform extractive and
Tris purification. The invention improves traditional guanidinium
isothiocyanate method, adopts the two steps of
chloroform extration after
cracking and
phenol-adding for purification, and respectively extract high-quality
total RNA from the
leaf blade and seed case of red-skinned pears, the
leaf blade, fruit
flesh and seed case tissues of grapes, the fruit of strawberries and the
callus of
arnebia euchroma. The extraction buffer liquid in the method contains guanidinium
isothiocyanate,
sodium dodecyl sarcosinate,
sodium citrate, soluble
polyvinylpyrrolidone and beta-mercaptoethanol, which can not only crack
plant cells efficiently and release
RNA, but also effectivelyinhibit the activity of
RNA enzyme, and can also prevent the oxidation of phenolic compounds and suppress the interference of secondary metabolites. In the second step,
Tris purification is utilized for crude extraction of RNA solution, which can eliminate the polysaccharides and
protein which can coprecipitate with RNA. The invention is simple in operating steps, economic and practical, good in stability, and suitable for extracting high-quality
total RNA from
plant tissue rich in polysaccharides and polyphenols and secondary metabolites.