GSM7989398: N2 young adults, 25oC, rep4; Caenorhabditis elegans; RNA-Seq
Frozen worm aliquots were thawed, 300 µL of TRIzol (Life Technologies, 15596026) were added and mixed vigorously. Six freeze-thaw cycles were employed to dissolve animals, specifically by freezing tubes in liquid nitrogen for approximately 30 seconds, and subsequent thawing for approximately 3 minutes. After each cycle, tubes were mixed vigorously. After the sixth freeze-thaw cycle, samples were spun down at 20,000 G for 3 minutes and the supernatant was subsequently collected into a fresh tube. Next, 1 volume of 100% ethanol was added to the sample and mixed vigorously. Finally, the mixtures were loaded onto Direct-zol columns (Zymo Research, R2060) and manufacturer's instructions were followed. In-column DNase I treatment was included. Quality control of samples, library preparation (non-directional, with poly-A enrichment), and mRNA sequencing (Illumina, PE150) was performed by Novogene.