Generic sample from Caenorhabditis elegans
Caenorhabditis elegans
 For the lin-35(n745) L1s, we grow the strain at 20 deg until majority of animals are gravid.  2) We bleach the worms to collect embryos, then place the embryos in S-basal buffer and incubate at 20 deg overnight to hatch into L1s.  3) We then put the hatched ~7K L1s on each of 5-6 seeded NGM plates at 20 deg, grow ~3 days until gravid again 4) We then bleach the worms for embryos, put the embryos in S-basal and incubate at 20 deg overnight to hatch into starved L1s. 5) We then place all L1s onto one large seeded plate for 4 hours at 25 deg.  6) We then wash off all worms from food and let them digest their ingested food for one hour in S basal 7) We spin down, monitor pellet size, and then add four volumes of Trizol, perform three rounds of freeze/thaw, and submit for RNA isolation. Run 30KCMAAXX = 150-250 bp size selection, 1% agarose gel, GAPipeline-1.0 Run 30k3FAAXX = 250-350 bp size selection, 1% agarose gel, GAPipeline-1.0 Run 311T6AAXX, 311U1AAXX = 250-350 bp size selection, 1.5% agarose gel, GAPipeline-1.3 Quality values = solexa/illumina  (not converted to phred) 