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文献和实验Purification of Plasmid from 50 ml-culture
at room temperature in a plastic bottle. Don''t autoclave. Solution III 5M potassium acetate 60 ml glacial acetic acid 11.5 ml Distilled water 28.5 ml Store at room temperature. This solution can be autoclaved. However, we usually
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
Carnoy's Fix: 6 parts 100 % EtOH 3 parts CHCl3 1 part glacial acetic acid Carmine Alum Stain: Place 1 g carmine (Sigma C1022) and 2.5 g aluminum potassium sulfate (Sigma A7167) in 500 ml distilled water and boil for 20 min. Adjust final
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