摘要:Adapted from Yaffe,M.P.and Schatz,G.,PNAS,1984,81,4819-4823.1.Grow 25mls yeast cells to 5x10E6.2.Sequentially spin down cells in a 15 ml polypro tube.3.Wash cells with 1ml ice cold ddH2O,transferrring[阅读全文:]
摘要:Horvath and Riezman,Yeast,1994; Gottschling LabSample Buffer: 10 ml 0.06M Tris-HCl,pH 6.8 0.6 ml 1M Tris 6.8 10% (v/v)glycerol 2 ml 50% glycerol 2% (w/v)SDS 2 ml 10% SDS 5% (v/v)2-mercaptoethanol 0.5 [阅读全文:]
摘要:To concentrate proteins for analysis by SDS PAGE:If a small amount of protein is to be precipitated (less than a few micrograms),add Insulin as a carrier protein (10 micrograms of Sigma insulin,I-5500[阅读全文:]
摘要:SummaryWCE Extracts are prepared from strain SHY282 (TFC4-Flag-EE (HpaI)).TFIIIC complex is first purified by flag affinity: TFC4 contains a single copy of the flag epitope.The flag purified TFIIIC is[阅读全文:]
摘要:Purification of TFIIA from E.coliLinda Warfield,Hahn Lab,2003Volumes given are for 2L of cells for each subunit (Toa1 and Toa2).Toa1 and Toa2 subunits are expressed separately in cultures of OD0.6 for[阅读全文:]
摘要:点击下载PDF文档[阅读全文:]
摘要:点击下载PDF文档[阅读全文:]
摘要:Buffer A Buffer B50 mM tris-HCl, pH 8.0 20 mM Na2HPO4, pH 7.25 mM EDTA20 mM NaCl10 mM NaCl5 mM EDTA25% (w/v) sucrose[阅读全文:]
摘要:Buffers:Lysis Buffer (1 liter)50 mM NaH2PO4 6.90g NaH2PO4.H2O (MW 137.99g/mol )300 mM NaCl(up to 2M)17.54gNaCl(MW 58.44)or 60ml 5M10 mM Imidazole (up to 100mM)0.68g (MW 68.08 )10 mM BME (up to 20 mM)0[阅读全文:]