title: GSM1470302: EB_activin-_shHEB_rep3; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was isolated using the RNeasy Plus Mini Kit (Qiagen), and messenger RNA (mRNA) was purified from the total RNA using Dynabeads Oligo(dT)25 (Invitrogen) according to the manufacturer's instructions. RNA sequencing libraries were built using an approach which targets the 3′ end (3SEQ) as described previously (Beck et al., 2010). Heat sheared mRNA was reverse transcribed with the P7_oligo-dT Index primers. The P5 linker was then ligated to the free end, and the linker-ligated cDNA was size-selected for 200-350 bp fragments by 3% Nusieve agarose gels (Lonza). The selected DNA was amplified using primers P5 and P7_Index for 15 cycles, and the amplified product was second size-selected for 200-350 fragments. The purified library quality was assessed using Bioanalyzer and Qubit, and sequenced on the Illumina HiSeq 2000 with multiplexing.