RNA Extraction from Purified Virus Particles

Abstract

The extraction procedure involves three consecutive extractions with phenol and three washings of the extracted RNA with 70 % ethanol. Several measures ensure high quality of the RNA:

• Working under conditions of high pH in which the binding between RNA and coat proteins is weakened

• Applying macaloid (a clay) to adsorb RNase

• Use of SDS to denature RNase

• Adding EDTA for binding oxidising ions, such as Fe^3+, which may otherwise generate harmful oxygen radicals

• Use of redistilled phenol to avoid the presence of oxidised polyphenols

• Use of chloroform to remove fatty acids

• Use of isoamyl alcohol to avoid generation of foam due to the presence of SDS