发布时间:2019-08-20 16:14 原文链接: YeastLysatesforWesterns

  • Cells are grown for 2-3 days as 1.5ml prep. under selection for the plasmid of interest. Spin cells down 2.6K for 5min.

  • Resuspend in 1ml 0.25m NaOH/1% 2-mercaptoethanol

  • Incubate 10min on ice.

  • Add 0.16ml 50% trichloroacetic acid

  • Incubate for 10min on ice.

  • Pellet 14K 10min.

  • Resuspend pellet in 1ml ice cold acetone.

  • Pellet 14K 10min.

  • Air dry pellet and resuspend in 200-500ul SDS SB and proceed as described below in section on western transfer