Two days after transient transfection, 293T cells were harvested and washed with ice-cold phosphate-buffered saline before being disrupted with lysis buffer (50 mM Tris, 100 mM NaCl, 0.1% NP-40, 50 mM NaF, 1 mM dithiothreitol, 1 mM phenylmethylsulfonyl fluoride, 1 μg/ml of aprotinin, 0.5 μg/ml of leupeptin, and 0.7 μg/ml of pepstatin).
After adding the fragmentation buffer, mRNA was disrupted into short fragments (≈200 bp).
Cells (5 × 10) were swollen in ice-cold hypotonic CaRSB buffer and were disrupted using a Dounce homogenizer.
The parasites were resuspended in R buffer at 5×108 parasites/ml and were disrupted in the French Press as described.
The pellets were suspended in Tris buffer and the cell walls were disrupted in a Microfluidizer M-110L.
The cells were lysed in RIPA buffer [ 32], and DNA was disrupted by sonification (LKB Instruments).
After suspended in Tris HCl buffer (50 mM, pH7.5), cells were disrupted by sonication.
Briefly, after adding formaldehyde, A549 cells were suspended in SDS lysis buffer and the chromatin DNA was disrupted by sonication.
The collected cells were suspended in Buffer A (20 mmol/L potassium phosphate, 100 mmol/L NaCl, 0.1 mmol/L EDTA, 10 mmol/L 2-mercaptoethanol, 5% glycerol, pH7.0) containing 1 mmol/L PMSF before the cells were disrupted by sonication.
The breakpoint was defined as the last ratio completed before the response was disrupted.
