GSM8493758: human primary PTEC, PBS control, biol_rep4; Homo sapiens; RNA-Seq
At the conclusion of 6h incubation, cells were lysed, harvested, and RNA was extracted using the Qiagen RNAEasy kit according to instructions for adherent animal cells. RNA-seq libraries were prepared using the TruSeq Stranded mRNA protocol (Illumina). Total RNA integrity was checked on a Bioanalyzer NanoChip (Agilent). Poly(A)+ RNA was then selected using oligo-d(T)-coated magnetic beads, followed by fragmentation using divalent cations and heat. Fragment RNA was converted to double strand cDNA using dUTP in the second strand synthesis to enforce strandedness in the final library. Indexed adapters were ligated to the cDNA and the library was then amplified by limited rounds of PCR. The final libraries were profiled on a Tapestation D1000 tape (Agilent) and quantified by real time PCR (Kapa Biosystems) on a StepOnePlus real time PCR workstation (Thermo/ABI).