Purification of calf thymus RNA polymerase II for in vitro transcription studies.
A procedure for the purification of a large amount of RNA polymerase II (RNAP II) from calf thymus is described. This procedure results in an approximately 1400-fold purification with 40% yield of enzyme in 60 h. The partially purified enzyme is highly suitable for in vitro transcription studies in a cell-free system utilizing HeLa S-100. This method for RNAP II purification would find significant applications in in vitro studies for the analysis of factors modulating eukaryotic transcription.
Laboratory of Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
J S
JS
Tyagi
I
I
Pastan
D000818:Animals; D002417:Cattle; D002474:Cell-Free System; D002642:Chick Embryo; D004795:Enzyme Stability; D012319:RNA Polymerase II / Q000302:isolation & purification*; D013698:Templates, Genetic; D013950:Thymus Gland / Q000201:enzymology*; D014158:Transcription, Genetic
D012319:RNA Polymerase II