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文献和实验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
is an indirect expression of the monoclonal antibody amount in the sample.Methods1 Obtain a 96-well plate and rinse 3x with PBS.2 Add 50 µl of a 1 mg/ml of antigen (venom, serum, or fraction), incubate for 30 min at 37°C.3 Rinse with PBS 3x.4 Add 5% powdered milk
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
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