Extraction of RNA from Yeast Packed into Column without Isomerization

Abstract

An RNA preparation containing only. 3'-5' phosphodiester linkages was investigated. Such RNA was extracted from yeast packed together with a filter aid (Celite 545) into a column with diluted alkali-salt solution in temperature below 40°C. The RNA from the cells immediately neutralized the surrounding alkali, so that a neutral solution of RNA was obtained as extract. The RNA yield was higher by column extraction than by usual flask extraction with hot salt solution. The enzymatic degradation of RNA during column extraction was prevented by preheating the yeast at 100°C for 2 hr or by extraction in temperature below 0°C, in the presence of high concentration of salt.