Purification of DNA-dependent RNA polymerase II from Saccharomyces cerevisiae.
A rapid procedure for the purification of RNA polymerase II from Saccharomyces cerevisiae is described. Total RNA polymerase activity was solubilized from whole cells by sonication in 0.32 M (NH4)2SO4 and RNA polymerase II purified by polyethylenimine fractionation, ammonium sulfate precipitation, and chromatography on DEAE-cellulose, DEAE-Sephadex, and phosphocellulose. The procedure may be completed in 2.5 days and the resultant enzyme is judged to be greater than 90% pure.
D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange; D012321:DNA-Directed RNA Polymerases / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D012319:RNA Polymerase II / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
D012319:RNA Polymerase II; D012321:DNA-Directed RNA Polymerases