GSM5200845: HEMnDP2 control2; Homo sapiens; RNA-Seq
Cells were pelleted and lysed in triozol and 1/5 volume of chloroform was added to each sample. Phase separation was performed by centrifugation. Supernatant was recovered and an equal volume of ethanol was added. This solution was added to the QIAGEN RNeasy mini Kit column (QIAGEN) and subsequent steps were performed according the the manufacturers protocol, including on column DNase digestion. Libraries were constructed using 1ug of RNA with the NEBNext Ultra II RNA Library Prep kit (NEB) with Poly-A mRNA enrichment was performed using NEBNext Poly(A) enrichment (NEB) protocol according to the manufacturer's guidelines. Libraries were indexed with NEB NEXT i7 index primers (NEB). Libraries were pooled to 10 nM and spiked 10% with phiX. They were sequenced 75 bases single-end on Illumina HiSeq 2500.