RNA-seq of HUVEC cells treated with Media (Water): Sample FUNGEI1-HT42
"Treated cells were collected by centrifugation at 2000 rpm and washed 2x using ice cold PBS. Collected pellets were lysed on the plate, using Lysis/Binding Buffer (Ambion), and frozen at -80oC. Poly-adenylated mRNAs were subsequently isolated from thawed lysates using the Dynabeads mRNA Direct Kit (Ambion) and following the manufacturer instructions. RNA-seq libraries were prepared using a protocol modified from the NEBNext Ultradirectional (NEB) library preparation protocol to use 96 Barcodes from BIOOScientific added by ligation, as described in Moyerbrailean et al. (2015). The individual libraries were quantified using the KAPA real-time PCR system, following the manufacturer instructions and using a custom-made series of standards obtained from serial dilutions of the phi-X DNA (Illumina)."