GSM6280587: BY273 control day 1, replicate 1; Caenorhabditis elegans; RNA-Seq
Total RNA was isolated with RNeasy mini kit (Qiagen) following the manufacturer's instructions. Reverse transcription was performed using High-Capacity cDNA Reverse Transcription kit (Applied Biosystems). Quantitative PCR was carried out on a QuantStudio 7 Flex detection system (Applied Biosystems) with the Power SYBR green PCR master mix (Applied Biosystems). Each sample was analysed in triplicate. Primer sequences are provided in Supplementary Table. 2. Sequencing libraries were prepared from 200 ng total RNA using Nugen Universal Plus Total RNA-Seq library preparation kit (Tecan) with custom AnyDeplete design for C. elegans. Final libraries were pooled (10 samples/run) and paired-end sequencing (2 x 71 bp) performed using NextSeq High Output kits on the NextSeq 550 sequencer. The raw sequencing data were demultiplexed using BCL Convert (Illumina). Sequencing data preprocess starts right after being demultiplexed. Sequences were aligned to C.elegans WBcel235 dna reference (release-96)26 using annotations, general transfer format (.gtf) file from the same release, and with STAR aligner (version 2.7.10a)27.