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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
Combined 3C-ChIP-Cloning (6C) Assay: A Tool to Unravel Protein-Mediated Genome Architecture
analysis. Restriction enzyme buffer (10X) Dilute the 10X stock to 1.14X for Steps 12 and 13. RNase A, DNase-free (10 mg/mL; Sigma R6513) SDS (Sodium dodecyl sulfate; 20% w/v; Fisher BP166) Sodium acetate (3 M, pH 5.2
water. Cool. Adjust pH to 7.0 with 1 M potassium hydroxide (about 35 mL) Dilute to 100 mL with more water. Washing solution: 25 mL 1.0 M potassium citrate buffer 50 mL 0.1 M magnesium sulfate heptahydrate (F.W. 246.5) 50 mL 0.1 M N-ethyl
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