The Larochelle Lab
at Clark University
Lab Protocols
RNA ISOLATION
Lyse 1x108 cells in 1 mL Trizol.
Lysate can be stored at -20ºC.
Spin lysate at 12000xg, 10 min, 4ºC.
Keep supernatant and incubate 5 minutes at RT.
Add 200 uL chloroform.
Vortex.
Incubate 3 min at RT.
Spin at 12000xg, 15 min, 4ºC.
Keep aqueous layer (top).
Add 0.5 mL isopropanol and incubate 10 minutes at RT.
Spin 12000xg, 10 minutes, 4ºC.
Wash pellet with 1 mL 75% EtOH (in DEPC-water). Vortex.
Spin 7500xg, 5 minutes, 4ºC.
Air dry 5 minutes.
Dissolve RNA in 500uL DEPC-water.
Incubate at 55ºC for 10 minutes.
Store at -80ºC.
Links | About Prof Larochelle
Clark Biology Dept | Clark University