Transcriptome sequencing O_mossambicus Tesis
RNA was extracted from tissue using TRIzol Reagent (Invitrogen), as per the manufacturer’s protocol. Following extraction, RNA was assessed for quality by visualisation on a 1.2% denaturing formaldehyde agarose gel, quantified using a Nanodrop spectrophotometer (Thermo scientific). For complimentary DNA (cDNA) synthesis, RNA samples were subjected to oligo-dT selection for mRNA purification and fragmented into small fragments. Fragmented RNA samples were subsequently repaired before adapter ligation. Suitable fragments were selected and reversed-transcribed into double-stranded cDNAs. The cDNA libraries were constructed by PCR amplification using random hexamer primed cDNAs. Finally, the samples were used for library construction and sequenced using an Illumina HiSeq 2000 sequencing (BGI, Hong Kong).