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But then came the second treadmill session: five minutes at maximum incline.
I like to spend ten minutes at maximum getting my face ready for the day.
At that time, samples were thawed and centrifuged for five minutes at maximum speed.
The ganglion was homogenized thoroughly with a pestle and the solution was centrifuged for 5 minutes at maximum speed.
The cell lysate was centrifuged for 5 minutes at maximum speed before transferring the supernatant to a new reaction vial.
The final pellet was re-suspended in 100 µl sterile PBS and centrifuged for 2 minutes at maximum g force, and the supernatant was sterile filtered.
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The supernatant was washed repeatedly with 1 ml equilibrated phenol (pH 7.5) until no more colouring was removed, each time vortexing for 5 seconds and separating both phases by centrifuging for 1 minute at maximum speed (20,000 × g).
Acaryochloris strains HICR111A and CRS proved very difficult to keep in suspension and were therefore sonicated (Misonix sonicator 4000, Qsonica LLC., Newtown, CT, USA) for one minute at maximum speed prior to spectrophotometric measurements.
DNA pellets were resuspended in 10 mM Tris-HCl, pH = 8 containing 10 μg/ml RNase A (Qiagen) and mixed for 1 minute at maximum speed on a multi-tube vortexer.
300 μl of Binding buffer PB was added to the coupled cDNA, and the mixture applied to a MinElute column (Qiagen, Valencia, CA), centrifuged for 1 minute at maximum speed and washed twice with 400 μl of buffer PE.
The solution was incubated for a further 10 minutes, spun at maximum speed for 30 s and the supernatant was discarded.
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