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文献和实验Replication timing by density transfer
Calbiochem's Pronase (catalog #53702), which seems pretty clean--i.e., plates spread after Pronase treatment aren't contaminated--but I heard from someone who'd used Sigma's Pronase and had massive growth of Streptomyces. 20x SCP = 0.6 M Na2 HPO
Blue Native Gel Electrophoresis
Blue Native Gel Electrophoresis Stock solutions 49.5%T, 3%C Acrylamide 24 g acrylamide, 0.75 g bisacrylamide / 50 ml H2O Store at RT 3 x Gel buffer 150 mM BisTris-HCl, 1.5 M 6-amino-caproic acid, pH 7.0 Adjust pH to 7.0 with HCl at 4°C
FINGERPRINTING PROTOCOL (AGAROSE GEL)
. Pour into 20X25 cm UV transparent trays. Insert comb. Use 200 ml of molten agarose. After gel is solidified, remove comb and place in electrophoresis unit with enough 1X TAE buffer to cover the gel. Load 1.75 µl of the restriction enzyme digestion
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