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文献和实验Recipe for yeast tryptone medium
. coli growth Yeast tryptone broth/agar, 2xYT Component stock conc final conc. /1l Tryptone - - 16g Yeast extract - - 10g NaCl - - 5g for agar plates agar - - 15g autoclave. Recipe for ammonium acetate
Preparation of cell lysates from E. coli by enzymatic lysis
(phenylmethylsulfonyl fluoride)in isopropanol 1M MgCl2Procedure1.Resuspend the cells in chilled lysis buffer in a ratio of 1 g cell wet weight to 1 ml lysis buffer.Add the PMSF (10 µl PMSF (100 mM)per ml of celsuspension)at this point.2.Add lysosyme to a final
Yeast Nuclear Extract (Small Scale)
50 ml glycerol adjust pH with KOH 3 mM DTT and protease inhibitors are added just before use Buffer C+ 75 mM Ammonium sulfate (1.5 liters) 20 mM HEPES 7.6 7.14g HEPES 10 mM MgSO4 15 ml 1M MgSO4 1 mM EGTA 6 ml 0.25 M EGTA 20% glycerol 300 ml
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