set-6_ok2195_Young
Worms were synchronously cultured on NGM plates, and transferred to NGM plates containing 20 M FUDR. Worms were harvested at day 2 of adulthood, washed several times using 14 buffer, and then lysed with the TRIzol reagent (Invitrogen). Total RNA was extred using the RNeasy Mini Kit (QIAGEN). The RNA quality was assessed by the Agilent 2100 and samples with RNA intensity (RIN) above 8.0 were used for construction of the library. Then, mRNAs were purified with Oligo(dT) magnetic beads and cDNAs were synthesized using Oligo (dT) primers. The bead-bound cDNAs were subsequently digested with the restriction enzyme Nla and ligated with the Illumina adaptor 1 at the sticky 5' end. Then, the bead-bound cDNA fragments were digested with Mme I and ligated with Illumina adaptor 2 at the 3' ends. After linear PCR amplification, products were purified by PAGE Gel electrophoresis.