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文献和实验M199 + 35g NaHCO3 + 25mM Hepes + 5 mM glutamine + 50 ug/ml Gentamycin.1M Hepes: 119.1 g Hepes + 500 ml dH2 O.Media A: 1X HBSS (10X:50ml) + 10 mM Hepes (1M: 5 ml) + 5 mM EDTA (0.5M: 5ml) + 2.5 % FBS (12.5 ml) in 500 ml.Media B: RPMI1640 470 ml + Gent
-Cl, pH8.0 1.2114 20% (v/v) Glycerol 20g 4% (w/v) SDS 4g 0.2% Bromophenol blue 0.2g 200mM DTT 20ul 5x Running buffer: 1 L 15.1 g Trizma base (= 125m M) 94g Glycine (= 1.25 M) 50ml 10%SDS (= 0.5%)--add last Do not adjust the pH!! 10x Blotting
A quick RNA mini-prep for Neurospora mycelial cultures
is as follows: 1. Conidia from slants (grown in 16 x 150 mm test tubes containing 8 ml of solid medium) are resuspended in 50 ml of Horowitz complete medium (Horowitz 1947 J. Biol. Chem. 171:255-262) and the cultures grown overnight with shaking at 30°C
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