title: GSM8218448: CD4+ T cells, IL-2 added, 96h, biol rep 1; Mus musculus; RNA-Seq
library_construction_protocol: RNA was produced using the NucleoSpin® RNA, MACHEREY-NAGEL kit. 300-400ng of total RNA was used for the construction of sequencing libraries. 33 Libraries were prepared using the INCPM-mRNA-seq. Briefly, the polyA fraction (mRNA) was purified from 300-400 ng of total input RNA followed by fragmentation and the generation of double-stranded cDNA. After Agencourt Ampure XP beads cleanup (Beckman Coulter), end repair, A base addition, adapter ligation and PCR amplification steps were performed. Libraries were quantified by Qubit (Thermo fisher scientific) and TapeStation (Agilent). Samples were sequenced on 4 lanes of Illumina NextSeq 500. Sequencing was done on a Nextseq 75 cycles high output kit, allocating 15M reads per sample (Illumina; single read sequencing).