TRIzol method for RNA isolation


 * 1) chunk one N2 (or mutant) 100mm plate to 4-6 100mm plates seeded with OP50
 * 2) let grow at 20° (unless doing a prep on a temperature sensitive mutant) until the plate is dense and most of the OP50 is gone (most of the animals should be adults and L4s); this should take 2-4 days depending on the density of the plate that was chunked
 * 3) wash worms off these plates with M9 media into a 1.5ml eppendorf tube (you may need to spin the tube down several times to get all the washes in one tube)
 * 4) after getting all the worms in one tube, wash 1X with 1ml of M9 media
 * 5) add 800ul TRIzol
 * 6) homogenize for 60 sec (by vortexing) - add 160ul chloroform (1/5 volume of Trizol)
 * 7) mix by vortexing for 15 sec
 * 8) incubate for 5 min at room temperature
 * 9) centrifuge at 12,000 g for 15 min at 4°C
 * 10) transfer the aqueous phase into a new tube
 * 11) add 500ul isopropanol
 * 12) centrifuge at max for 10 min in the cold room
 * 13) wash the pellet with 500ul 70 % ethanol
 * 14) centrifuge at max for 2 min in the cold room
 * 15) air-dry the pellet for 5-10 min - disolve the pellet in 50-100 ml DEPC-H2O
 * 16) OD prep at 260nm, you’ll need 1ug for RT-PCR (prepping 2 100mm plates of N2 worms typically give total RNA at a concentration of .5ug/ul)