title: GSM8992589: Differentiated LUHMES with 4sU and HSV-1 12hpi Rep2; Homo sapiens; Human alphaherpesvirus 1; RNA-Seq
library_construction_protocol: Cells were lysed in TRIzol and RNA col-lected with the Direct-zol RNA Miniprep kit (Zymo Research #R2050). Total RNA was treated with DNase I and then Ribo-Zero before fragmentation to 130-160nt.  cDNA synthesis was combined with End Repair Mix, then add A-Tailing Mix,  then the Adenylate 3'Ends DNA, RNA Index Adapter and Ligation Mix before  Ampure XP Beads purification. The double stranded PCR products were heat denatured and circularized by the splint oligo sequence. The single strand circle DNA (ssCir DNA) were formatted as the final library. The library was amplified with phi29 to make DNA nanoball (DNB) which have more than 300 copies of one molecular. The DNBs were load into the patterned nanoarray and paired-end 100 base reads were generated in the way of combinatorial Probe-Anchor Synthesis (cPAS).