Trizol extraction


 * Move TRIzol bottle from 4º to rt
 * Add 520µl TRIzol to cells and lyse in plate for 1 min
 * Transfer 500µl lysate to tubes
 * Add 100µl chloroform and vortex for 10 sec.
 * Incubate at rt for 2 min
 * Spin 10,000g for 10 min at rt
 * Take aqueous layer (top layer) and transfer to new tube (~200µl)
 * Add equal amount of isopropanol (~200µl)
 * Incubate at rt for 20 min
 * Spin 10 min at 4º max speed
 * Wash pellet with 70% EtOH
 * Spin 5 min at 4º max speed
 * Carefully aspirate supernatant (remove last trace with pipet)
 * Dry pellet
 * Resuspend RNA in RNase-free water (~30µl)
 * warm 5 mins @ 65C to help re-suspend RNA pellet

Note: scale up proportionally. Can also scrap cells in trizol before transferring. Can freeze in trizol (-80), or precipitate O/N @-20 (iso step), or can freeze RNA pellet (very stable).