title: Illumina HiSeq 1500 sequencing; Small RNA and total RNA sequencing of C. elegans with mutations in the hrde-1 and emb-4 genes
design: Small RNA and total RNA sequencing of C. elegans with mutations in the hrde-1 and emb-4 genes
library_construction_protocol: Animals were harvested and washed 3X in M9 buffer. Synchronised animals were grown to young adult stage at 20C on HB101 seeded NGM plates. Settled animals were mixed with Trisure reagent, bead beaten using zirconia beads, and the RNA is extracted using chloroform. Small RNA sequencing is performed by treating 5ug of total RNA with Epicentre 5’ polyphosphatase to remove the 5’ triphosphate. After treatment, RNA is purified by phenol/chloroform extraction and 1ug of RNA is used to prepare small RNA libraries using Illumina TruSeq small RNA library preparation kit.