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文献和实验TOP10 chemically competent cells
10 mM KOAc pH 7.0 (10 ml of a 1M stock/L) 80 mM CaCl2 .2H2 O (11.8 g/L) 20 mM MnCl2 .4H2 O (4.0 g/L) 10 mM MgCl2 .6H2 O (2.0 g/L) 10% glycerol (100 ml/L) adjust pH DOWN to 6.4 with 0.1N HCl if necessary
Fast and reliable mini-prep RNA extraction from Neurospora crassa
otherwise. - Pulverize the mycelium in a mortar with liquid nitrogen - Transfer the powder into a mixture of 0.75 ml lysis buffer (0.6 M NaCl, 10 mM EDTA, 100 mM Tris HCl, pH 8.0, 4 SDS) and 0.75 ml phenol (saturated with 0.1 M Tris HCl, pH 8.0) in a 2 ml
or cosmid DNA of interest into a 12 X 75 mm Falcon tube containing 2 ml of LB media supplemented with the appropriate antibiotic (typically ampicillin at 100 ug/ml) and incubate at 37deg C 8-10 hours with shaking at 250 rpm. Transfer the culture
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