GSM7747687: TdLN 2; Mus musculus; RNA-Seq
Tumor and lymph nodes were excised, digested and lymphatic endothelial cells were FACS-sorted based on CD45negCD31+gp38+. They were collected in RNAprotect Cell Reagent (Qiagen). RNA was isolated using RNeasy Plus Micro kit (Qiagen) cDNA libraries were constructed by the Genomic platform of the University of Geneva as followed: the SMARTer™ Ultra Low RNA kit from Clontech was used for the reverse transcription and cDNA amplification according to manufacturer's specifications, starting with 1 ng of total RNA. 200 pg of cDNA were used for library preparation using the Nextera XT kit from Illumina.Pools of 10 libraries were loaded for clustering on a Single-read Illumina Flow cell. Reads of 50 bases were generated using the TruSeq SBS chemistry on an Illumina HiSeq 4000 sequencer.