For luciferase measurement, sample wells were washed twice with phosphate-buffered saline, followed by addition of cell lysis buffer (Promega, Madison, WI); the plates were then shaken for 20 min at room temperature to allow cell lysis.
The resulting loaded PEG-liposomes were then shaken.
The cultured cells were then shaked thoroughly for 1 minute.
The mixture was then shaken by hand.
This plate was then shaken at 37°C for 16 hours.
The extract was then shaken for another hour.
50 µl of beads (in binding/wash buffer) were then added to RNA, incubated at 42°C for 20 min, and then shaken for 2 h at room temperature.
Worms in L1 diapause suspended in liquid buffer were then exposed to an acute heat stress at 36.8° in a shaking incubator (70 rpm) for 4 hr in a sealed microcentrifuge tube.
Chelation buffer was then removed and 5 ml of shaking buffer (PBS with 43.3 mM sucrose and 54.9 mM sorbitol) was added.
The reaction buffer was then changed to a fresh one and the gels were incubated (24 h, 37°C) in a shaking incubator.
