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文献和实验Replication timing by density transfer
. (I add 0.1 mg/ml ampicillin if I'm feeling paranoid.) The doubling time of RM14-3a is ~ 2.6 hr at 23o C in glucose and 6-8 hr (depending on aeration) in acetate--longer if you're selecting for a plasmid. RM14-3a is MATa, cdc7-1ts , bar1, trp1-289
Saturation Analysis of Ligand Binding Using a Centrifugation Procedure
) HTS‐EDTA: 5 mM HEPES/4.5 mM Tris⋅Cl /1 mM EDTA, pH 7.4 (pH adjusted, if necessary, with concentrated HEPES‐free acid or Tris‐free base), ice cold Unlabeled ligand
, 10 μg/mL leupeptin, 1 mM phenylmethanesulfonylfluoride (PMSF), and 1 mM sodium orthovanadate. 6. Teflon cell scrapers (Fisher, Pittsburgh, PA ). 7. 2× SDS sample buffer for electrophoresis: 0.0375 M Tris-HCl, pH 6.5, 8% (w/v) SDS, 10% (v/v
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