title: 66K00384,PBMC,batch 1,time 1
design: 500 ng total RNA was used as input; adapter-ligated library prepared with the KAPA HyperPrep Kit using Bioo Scientific NEXTflex DNA Barcoded; rRNA depleted by incubation with complementary probes; RNase H was used to degrade rRNA; cleanup and DNase digestion with Kapa Pure Beads and DNase; fragmention at 85 ºC for 4.5 minutes in the presence of magnesium; NEXTflex DNA Barcoded Adapters were ligated to A-tailed cDNA; products purified with Kapa Pure Beads; 8 cycles of amplification; cluster formation on Illumina cBOT; sequencing to a target of 40–50 million 2x126 bp paired-end reads.