title: GSM9460347: CF2166, 72h, sRNA, rep1; Caenorhabditis elegans; ncRNA-Seq
library_construction_protocol: Worms were washed from plates and rinsed with M9 3 times and flash frozed in liquid nitrogen. Total RNA from cell lysates was isolated using trizol/chloroform and isopropanol precipitation. For small RNA libraries, small RNAs (~16-30-nt) were purified from total RNA by size selection using 17% polyacrylamide gels and treated with polyphosphatase 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 libraries, total RNA was DNase treated and rRNA was depleted using the Ribo-Zero rRNA Removal Kit (Human/Mouse/Rat, Illumina). rRNA-depleted RNA was size selected (>200 nucleotides) to remove 5S rRNA and tRNA using the Clean & Concentrator-5 Kit (Zymo Research). RNA-seq libraries were prepared using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB).