PURIFICATION USING POLYETHYLENIMINE PRECIPITATION AND LOW-MOLECULAR WEIGHT SUBUNIT ANALYSES OF CALF THYMUS AND WHEAT-GERM DNA-DEPENDENT RNA POLYMERASE-II
PURIFICATION USING POLYETHYLENIMINE PRECIPITATION AND LOW-MOLECULAR WEIGHT SUBUNIT ANALYSES OF CALF THYMUS AND WHEAT-GERM DNA-DEPENDENT RNA POLYMERASE-II
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DOI：
10.1021/bi00630a005
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发表时间：
1977-01-01
期刊：
影响因子：
2.9
通讯作者：
BLATTI, SP
中科院分区：
文献类型：
--
作者：
HODO, HG;BLATTI, SP
材料与方法将来自General米尔斯的小麦胚芽储存在4 ℃.新鲜冷冻的小牛胸腺从爱荷华州迪比克的迪比克公园获得，并保持在-70 ℃直至使用.所有生物化学品均为试剂级。氚标记的UTP（比活度，16 Ci/mmol）从施瓦茨/Mann获得，α-鹅膏蕈碱和二硫苏糖醇从Calbiochem购买，小牛胸腺DNA（I级）从Sigma Chemical Co.，牛血清白蛋白来自Miles实验室。聚乙烯亚胺（30%）得自Aldrich Chemical Co. Nonidet-P-40是Shell Chemical Co. Solutions的赠品。所有缓冲液均由反渗透纯化去离子水制备。将二硫苏糖醇制备为0.1 M储备溶液，并在使用前立即添加。缓冲
Materials and MethodsMaterials. Wheat germ from General Mills was stored at 4 C. Fresh frozen calf thymus was obtained from Dubuque Park, Dubuque, Iowa, and maintained at—70 C until use. All biochemicals were reagent grade. Tritium labeled UTP (specific activity, 16 Ci/mmol) was obtained from Schwarz/Mann, a-amanitin and dithiothreitol were purchased from Calbiochem, calf thymus DNA (grade I) was from Sigma Chemical Co., and bovine serum albumin was from Miles Laboratories. Polyethylenimine (30%) was obtained from Aldrich Chemical Co. Nonidet-P-40 was a gift of Shell Chemical Co.Solutions. All buffers were prepared from reverse-osmosis purified deionized water. Dithiothreitol was prepared as a 0.1 M stock solution and was added immediately before use. Buffer