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文献和实验and centrifuge 4 minutes at 2000 RPM;Remove the PBS+1/1000 PMSF by aspiration and re-suspend cell pellet in 1ml ChIP lysis buffer plus the protease inhibitors 1/1000 PMSF and 50 units of RNAse inhibitor. The final volume of cell lysis buffer should be sufficient
Replication timing by density transfer
volume of a 2x enzyme mix that contains the appropriate restriction enzyme buffer, restriction enzyme (usually 1 microliter per sample) and 0.25 microgram/ml RNase A. If the DNA prep is clean, the digest can go overnight at 37 degrees C.] Choosing
A quick RNA mini-prep for Neurospora mycelial cultures
temperature. [E buffer: 50 mM Tris-Cl pH 8.0, 300 mM NaCl, 5 mM EDTA, pH 8.0, 2 SDS; autoclave and add 1 mM ATA and 14 mM ß-mercaptoethanol. ATA=aurintricarboxylic acid, ammonium salt (Sigma #A0885, St. Louis, MO)] 5. Thaw the powder in E buffer in 42°C
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