GSM8482684: GFP::alg-1, nrfl-1 RNAi, co-IP, rep 1; Caenorhabditis elegans; RNA-Seq
Worms were washed from plates and rinsed with M9 3 times and flash frozed in liquid nitrogen. RNA was isolated using trizol/chloroform and isopropanol precipitation. Small RNAs (~16-30-nt) were purified from total RNA by size selection using 17% polyacrylamide gels and in some instances treated with NEB RNA 5' Pyrophosphohydrolase (RppH)to facilitate capture of 22G-RNAs. Libraries were then prepared according to the NEBNext Multiplex Small RNA kit protocol. PCR products (~134-148 bp) were size selected using 10% polyacrylamide gels. For mRNA-seq DNase-treated total RNA was depleted of rRNA using Ribo-Zero and libraries were prepared using the Illumina TruSeq Stranded Total RNA kit.