NextSeq 500 sequencing; RNA-seq of C. Elegans control vs SKN-1b mutant
RNA-seq of C. Elegans control vs SKN-1b mutant
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