title: GSM3754212: Donor 4 : CDC2 [AH27_4]; Homo sapiens; RNA-Seq
library_construction_protocol: Primary cell populations isolated by flow cytometry (n=200 cells per population) were captured directly into lysis buffer containing 0.1% triton X-100, 1U RNaseOUT RNase inhibitor, 2.5mM each dNTP mix, 2.5uM anchored oligo-dT primer (5'-AAGCAGTGGTATCAACGCAGAGTACT30VN-3') in a total volume of 20ul. Full-length mRNA was reverse transcribed and amplified using the Smart-seq2 protocol of Picelli et al (Nature Methods. 2013;10(11):1096-8). Sequencing libraries were prepared from 1ng cDNA product using Illumina Nextera XT reagents, in accordance with the supplier's protocol, except that PCR was limited to 8 cycles to minimize read duplicates, and final library elution was performed using 17.5μl elution buffer EB.  All libraries were pooled and sequenced on four lanes of an Illumina HiSeq 2500 sequencer, using a HiSeq Rapid Cluster Kit v2.
attributes.tag: GEO Accession
attributes.value: GSM3754212