After DNase treatment, aliquots of total RNA from all samples were individually was fragmented (300-500bp) and reverse-transcribed for first strand cDNA synthesis using oligo(dT)-index primer. And then a reaction pool per above 10 first strand cDNA libraries processed second strand synthesis, adapter ligation and finally amplified by 15 cycles of PCR using Q5 HiFi HotStart (NEB, E7645L) to be sequenced across multiple lanes on a NovaSeq 6000 machine.
