GSM6307608: WTC-CM 2D -Sample 3; Homo sapiens; RNA-Seq
GSM6307608
RNA was extracted with Trizol (Thermo Fisher) followed by a column based purification (Zymo Research). Eluates were stored at -80C prior to sequencing. One microliter of undiluted total RNA was incubated with 10ul of NEB Dynabeads for polyaA+ RNA selection, eluted, and fragmented in 12ul. Illumina libraries were constructed using NEB Ultra II protocol; 5ul of the fragmented solution using half the amount of reagent and 1,2 PrD instead of enhancer in the ligation step, with 16 PCR cycles of final amplification and duplex barcoding. Libraries were quantified using the Fragment Analyzer and qPCR before being sequenced on an Illumina NextSeq 500 using 75-bp paired-end reads.