RNA SEQUENCING OF OVCAR-3
The whole transcriptome library for sequencing is made using Sense Total RNA-seq Library prep kit . This protocol maintains the strand specificity to allow the mapping of the genes to the corresponding strand. The workflow involves depletion of ribosomal RNA, including the mitochondrial fraction, followed by the random hybridization of Starter/Stopper heterodimers containing Illumina-compatible linkersequences to the RNA template. Reverse transcription starts from the Starter oligo and ends at Stopper oligo where the cDNA is ligated. Then, the second strand is generated followed by PCR amplification where P5 and P7 adapters are added which is required for cluster generation