GSM3067518: YFP-PPARalpha-S280A1; Rattus norvegicus; RNA-Seq
Total RNA was isolated from H9C2 cells transduced with the indicated plasmid using TRIzol (Invitrogen).  Total RNA was subjected to two rounds of poly(A) selection using oligo-d(T)25 magnetic beads (New England Biolabs). A paired-end (strand specific) cDNA library was prepared using the NEB Next Ultra-directional RNA-Seq protocol. Briefly, poly(A)+ RNA was fragmented by heating at 94°C for 10 minutes followed by reverse transcription and second strand cDNA synthesis using the reagents provided in the NEB Next kit. End-repaired cDNA was then ligated with double stranded DNA adapters, followed by purification of ligated DNA with AmpureXP beads. cDNA was then amplified by PCR for 15 cycles with a universal forward primer and a reverse primer with bar code. 