RNA-Seq of Homo sapiens: peripheral venous blood
Total RNA underwent stringent quality control, followed by 3 and 5 adapter ligation.  Reverse transcription and PCR amplification were performed under optimized thermocycling conditions.  PCR products were purified via PAGE gel electrophoresis and eluted in EB buffer, and library quality was assessed using standard QC protocols.  Single-stranded DNA molecules were then generated by denaturation, circularized, and residual linear DNA was digested.  Circularized DNA molecules were amplified via rolling circle amplification to form DNA nanoballs (DNBs), which were loaded onto patterned nanoarrays and sequenced using combinatorial Probe-Anchor Synthesis (cPAS) technology.