title: GSM8127676: CD4+ T cell, IgG Control 3; Mus musculus; RNA-Seq
library_construction_protocol: RNA was extracted from lysates using the PureLink RNA Micro Scale Kit (Invitrogen) according to the manufacturer's instructions. 5ng of RNA was used for generation of sequencing libraries using the Quantseq 3' mRNA-seq Library Preparation kit (Lexogen). Briefly, library generation was commenced with oligodT priming containing the Illumina-specific Read 2 linker sequence. After first strand synthesis, RNA was degraded. Second strand synthesis was initiated by random priming and a DNA polymerase. Random primers contained the illumina-specific Read 1 linker sequence. Double stranded DNAwas purified from the reaction using magnetic beads and libraries amplified (18 cycles) and sequences required for cluster generation and sample indexes were introduced. Libraries were normalized and pooled at a concentration of 4nM for sequencing.