title: NextSeq 500 sequencing; RNA-seq of C. Elegans control vs SKN-1b mutant
design: RNA-seq of C. Elegans control vs SKN-1b mutant
library_construction_protocol: The worms were washed off from the plates using M9 and transfer to an RNase free tube. 500 µl of Trizol were added and to 10 cycles of freezing at -80°C, thaw at 37°C and vortex 30s were done. 125 µl of BCP (1-Bromo-3-chloropropane) were added, mixed by inversion and let for incubation 3 min at RT. Samples were centrifuged at 12000 rpm for 10 min at 4°C and the clear layer was transfer to a new tube. An equal volume of 70% ethanol was added and this was transfer to a column of the RNeasy Quiogen Kit. The skn-1b(tm4241) IV mutation is a deletion mutation of the skn-1b exon. So skn-1b is not expressed A RNeasy Quiogen Kit and the manufacturer's instructions were followed. The concentration and purity of the samples were checked using the nanodrop. PolyA selection kit