Purification of adenovirus messenger ribonucleic acid by an aqueous polymer two-phase system.
An aqueous polymer phase system containing 6.3% (w/w) dextran and 3.5% (w/w) poly(ethylene glycol) in 10 mM phosphate buffer (pH 8.0) was developed to select RNA-DNA hybrids from unhybridized RNA. The top phase of this phase system, which contains DNA and the RNA-DNA hybrids, can be used to purify adenovirus messenger RNA both early and late in the infectious cycle. The hybrids can be melted by heat in the top phase and the messenger RNA selected by oligo(dT)cellulose chromatography whereupon the polymers and the DNA percolate and the polyadenylated messenger RNA absorb to the column. The isolated messenger RNA appears to be almost quantitatively recovered at a purity from 70 to 90% depending on the concentration of the specific messenger RNA in the starting material. Early and late viral messenger RNA were selected on the complementary strands of adenovirus DNA according to this procedure.
D000260:Adenoviruses, Human; D003911:Dextrans; D006367:HeLa Cells; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008722:Methods; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D011092:Polyethylene Glycols; D012333:RNA, Messenger / Q000302:isolation & purification*; D012367:RNA, Viral / Q000302:isolation & purification*; D013696:Temperature
D003911:Dextrans; D012333:RNA, Messenger; D012367:RNA, Viral; D011092:Polyethylene Glycols