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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
ChIP Protocol-Mechanical Breakage & FA Lysis Buffer
One - Part One: 1. Grow culture until it is in log phase: OD600=0.7 - 1.2 2. For each sample you collect, put the appropriate amount of 37% formaldehyde in a 50ml tube and label. The calculation for the correct amount of 37% formaldehyde to use is: volume of culture/36
Hepes. Harvest cells by putting 20mls of -Ca-Mg PBS, buffered with 0.02M Hepes, and 1mM EDTA on each plate for 10min in the 37 C incubator. After the 10min, collect some of the PBS in a 50ml conical tube before pipeting
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