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文献和实验Blue Native Gel Electrophoresis
, I put the power supply on a timer when I change to dye-minus cathode buffer. The power supply turns off at the end of the run, but the cooling circulator remains on and the gel stays cold until the next morning, when it can be further processed. Immunoblotting
. 0.1% SDS. Keep a squirt bottle of it.VIII. Running buffer: Keep a jug of this. 24 g Tris base, 115.2 g glycine, 20 ml 20% SDS, H2O to 4 liters.IX. Sample buffer: 100 mM Tris, pH 6.8, 2% SDS, 5% ß- mercaptoethanol, 15% glycerol, enough bromophenol blue
with magnetic stand, and remove supernatent. d) wash once in 1X NEB TaqI buffer e) resuspend in 40 ul of 1X NEB TaqI buffer (w/1X BSA - 100X stock sent w/ enzyme ) Digestion: a) to 40 ul of cDNA add: 1 ul (10U) of Taq I. b) digest at 65C
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