GSM4263602: WT on control (RNAi) mRNAseq; Caenorhabditis elegans; RNA-Seq
Animals were washed off plates and washed twice with M9. Trizol was added to the worm pellet and frozen in liq N2. After thawing, the animals were ground using a motorized pestle fitting in an eppendorf tube, followed by Trizol extraction,  chloroform extraction and isopronanol precipitation. 18-28 nt RNA was size-selected on a 15% TBE-urea polyacrylamide gel, and eluted from gel. Small RNA was treated with RNA 5'polyphosphatase, followed by liagtion of the 3' adaptor. After hybridization of the Reverse Transcription primer, the 5' adaptor was ligated. cDNA was generated using Superscript III, and was amplified using Phusion polymerase.  The correct size product was isolated from a 10% TBE polyacrylamide gel. Total RNA libraries were prepared using epicentre Ribo-Zero magnetic kit, followed by use of the NEBNext Ultra RNA Library Prep Kit for Illumina2 kit. mRNAseq libraries were prepared using NEBNext Ultra magnetic isolation module and NEBNext Ultra Directional RNA Library Prep Kit
GEO Accession
GSM4263602