title: GSM3704451: WT3 LEC RNAseq; Mus musculus; RNA-Seq
library_construction_protocol: Lymphatic endothelial cells were isolated from lymph nodes by flow cytometry on the basis of CD45-, TER119-, CD31+, podoplanin38+ cells. Sorted cells were flash frozen on dry ice, and RNA was harvested using Quiagen RNA isolation kit. The library was sequenced on Illumina Hiseq 4000 sequencer as Single-Read 50 base reads following Illumina's instructions. Full length cDNA were generated from 4 ng of total RNA using Clontech SMART-Seq v4 Ultra Low Input RNA kit for Sequencing (Takara Bio) according to manufacturer's instructions with 10 cycles of PCR for cDNA amplification by Seq-Amp polymerase. 600 pg of pre-amplified cDNA were then used as input for Tn5 transposon tagmentation by the Nextera XT DNA Library Preparation Kit (Illumina) followed by 12 cycles of library amplification.