Method for extracting RNA from calcified tissue

Abstract

The invention discloses a method for extracting RNA from a calcified tissue. The method comprises the following steps: grinding the calcified tissue in liquid nitrogen until the blocky particle diameter is less than 0.5cm, adding Trizol, then adding the liquid nitrogen to continue to grind into powder until the diameter is about 1mm or less than 1mm; then transferring the obtained powder into a RNase-Free tubule, adding the Trizol for cell digestion lysis; centrifuging the tubule, transferring a supernatant to a new tubular; adding chloroform into the supernatant for treatment, then centrifuging, transferring an topmost layer aqueous phase to another new tubular, adding a 75% ethanol solution, mixing gently for nucleic acid precipitation; then transferring to an adsorption column in a kit, standing then centrifuging for RNA to bind to the adsorption column; using a kit solution for washing; and then eluting to obtain the total calcified tissue RNA. The obtained total calcified tissue RNA has the advantages of high purity, high yield, good quality and good integrity.

Description

technical field [0001] The invention relates to marine biological technology, in particular to a method for extracting RNA from calcified tissues, in particular to a method for extracting ribonucleic acid RNA from reef-building corals. Background technique [0002] Calcification is the process by which calcium salts deposit in soft tissue and harden it. The extraction of total RNA from calcified cells in calcified tissues is usually the first step in molecular biology experiments such as transcriptome sequencing, expression profile sequencing, cDNA cloning, RT-PCR and Northern blot of calcified tissues. The quality and quantity of total RNA determine Success or failure and quality of follow-up experiments. At present, there are many reports on the methods of extracting total RNA from cultured bone cell lines and various organs, but the method of extracting total RNA from calcified tissues has been in the groping stage. The purity and yield of RNA obtained by traditional ex...

AI Technical Summary

Problems solved by technology

[0008] Neither of these two methods can clearly know whether there is RNase residue in the RNA solution

If there is a very small amount of RNase in the solution, it is difficult to detect it with the above method, but most of the subsequent enzymatic reactions are performed above 37°C and for a long time

If there is a very small amount of RNase in the RNA solution, the RNase will have a very suitable environment and time to play their role in the subsequent experiment, which will lead to the failure of the experiment

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