To obtain nucleic acid, the basic steps include: cell lysis—nucleic acid extraction—nucleic acid washing—elution. After the cell lysis step, the RNA extraction can be performed either with organic solvent extraction or matrix adsorption method. Organic solvent extraction is a classic way to achieve high quality RNA. In the process of RNA extraction, chloroform is often used for extraction to remove impurities such as sucrose and protein, and to promote the separation of the aqueous phase and the organic phase, thereby achieving the purpose of purifying RNA. After extracting RNA by chloroform, isopropanol is generally used to precipitate aqueous phase RNA. After that, add 75% ethanol without RNase to resuspend the RNA pellet and sufficiently dissolve the salt ions in the RNA pellet. Then centrifuge again for 10-30 min to precipitate the RNA. After centrifugation, carefully dispose the supernatant, then add appropriate amount of RNase-Free deionized water to dissolve RNA precipitate.
In TIANGEN’s RNA extraction kits, TRNzol reagent and RNAsimple kit are based on this method, and total RNA with high yield, high purity and good integrity can be extracted within 1 hour.