GSM8186706: MOLT4.inh.4, rep 2; Homo sapiens; RNA-Seq
Follwing treatment, cells were immediately lysed in lysis buffer (from PerkinElmer's chemagic RNA Tissue 360 H96, VD200615) supplemented with BME flash frozen and stored at -80c. After all biological replicates were collected, samples were thawed in deep 96-well plates. An equal amount (~30pg) of Lexogen's ERCC spike-in mix (SIRV-3) were added at this point following manufacturer's recommendations. The PerkinElmer's Chemagic 360 was used for bead-based RNA extraction following manufacturer's standard protocol. RNA was quantified using an Agilent Fragment Analyzer, purified using 2X RNA SPRI beads, and 1ng of total RNA was used for Illumina library preparation. Libraries were prepared using a reduced volume version of SMART-seq2 performed on an SPT Mosquito HV using 10 cycles of amplification for cDNA generation and 16 cycles for library generation using NexteraXT (Illumina).