GSM5076858: Langerin+ cDC2_C; Homo sapiens; RNA-Seq
Primary cell populations isolated by flow cytometry were lysed and total RNA was extracted.  mRNA transcripts were reverse transcribed and amplified from 1ng total RNA, 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. Libraries were sequenced as pools over 1 or 2 lanes on an Illumina HiSeq 2500 sequencer.