GSM6422085: HeLa siTRESLIN 48 hours biol rep 1; Homo sapiens; RNA-Seq
Total RNA was extracted from cells using manufacturer's instructions (Cytiva Life Sciences). 1µg of total RNA was used for the construction of sequencing libraries. Libraries were constructed by BGI according to the following procedure (All reagents were made by BGI unless stated): mRNA molecules were purified from total RNA using oligo(dT)-attached magnetic beads, and fragmented into small pieces. cDNA was generated using random hexamer-primed reverse transcription, followed by a second-strand cDNA synthesis. The synthesized cDNA was subjected to end-repair and was 3' adenylated. Adapters were ligated to the ends of 3' adenylated cDNA fragments. cDNA fragments were PCR amplified and purified with Ampure XP Beads (Agencourt), and dissolved in EB solution. Library quality was validated on an Agilent Technologies 2100 bioanalyzer, and double stranded PCR products were heat denatured and circularized by the splint oligo sequence. Single strand circle DNA was amplified with phi29 to make DNA nanoballs (DNBs) with more than 300 copies of one molecule. DNBs were loaded into the patterned nanoarray and single end 50 basepair reads were generated on a BGISEQ-500.