GSM3290306: D7PrF_FMSt; Homo sapiens; RNA-Seq
Total RNA from 500K FACS-purified cells was extracted using RNEasy micro columns (Qiagen). RNA quality and quantity were measured by a TapeStation and NanoDrop. Total RNA was subjected to enrichment of polyA+ RNA using the NEBNext Poly(A) kit (NEB). Strand-specific polyA+ RNA-Seq was performed using the LM-Seq method, as described previously. Briefly, PolyA+ RNA was fragmented at 85°C in SmartScribe first-strand buffer (CloneTech) supplemented with a 3' sequencing adapter containing a random hexamer sequence. The fragmented RNA were subjected to first strand cDNA synthesis, followed by RNase A/H treatment. The remaining single-stranded DNA were ligated to a 5' sequencing adapter. The libraries were then amplified by PCR using Illumina TruSeq-compatible primers and purified using SPRI purification beads.
GEO
GSM3290306