Total RNA was prepared using TRIzol reagent (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol. Libraries with PolyA selection were constructed with the TruSeq Sample Prep Kit v2 (Illumina, San Diego, CA) following the manufacturer’s protocol. 2ug of total RNA was used for Poly-A selection and fragmentation, subsequently followed by cDNA systhesis, end repairing, dA tailing, adapter ligation and final library amplification. For “not-so-random” primer-based RNA-Seq library preparation, libraries were constructed following protocols described previously (Armour et al., 2009, Nature Methods). 1ug of total RNA was used for first-strand synthesis with Superscript III system (#18080044 Life Technologies, Grand Island, NY) and then performed second-strand synthesis with Klenow fragment (#M0212L, NEB). The final PCR amplification was performed with Expand High Fidelity Plus PCR system (#11732641001, Roche).
GEO Accession
GSM1829191