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文献和实验TRICINE GELS Protocol (For Low MW proteins (
Stock Solutions1) Gel Buffer: 3 M TrisHCl pH8.45 + 0.3% SDS. Keep RT. (Prepare: 18.15gr Tris + 150mg SDS + H2O up to 50ml. Adjust pH before you add the SDS) 2) 40% Acrylamide solution 29:1 (Acrylamide 29 / Methylene bis Acrylamide 1) or 40
[M]=A259 /15.4x103 =A259 1:2000 x0.130 adjust the final volume. Betaine monohydrate C5 H11 NO2 xH2 O; Mw=135.2g/M; (store at 4o C). Conc. Stock %(w/w) 100ml 200ml 700ml Betaine 135.2g/M 5M
Replication timing by density transfer
, leu2-3,112, ura3-52, his6 in an A364A background. I aim for about 32 x 106 cells per time point. That gives enough DNA for at least 3 blots, leaving enough leeway for errors. 2. When the cell density is 2 x 106 cells/ml (OD660 ~ 0.16
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