title: GSM7078021: 129_mRNA; Mus musculus; RNA-Seq
library_construction_protocol: For all samples, total RNA was extracted using Quick-RNA Microprep Kit (Zymo Research, R1050). 20ng of total RNA was used for each library. cDNA was generated from the RNA template using Template Switching RT Enzyme Mix (NEB, M0466) with hexamer priming. nanoCAGE-seq libraries were then prepared following the previously described protocol (Poulain, S. et al. Promoter Associated RNA, Methods and Protocols. Methods Mol Biology Clifton N J 1543, 57–109 (2017).). For each of the two B6 samples (Ly49D+ and Ly49D-), we also generated one additional library where we used 250ng total RNA as input, extracted mRNA, and further depleted truncated mRNA with Terminator™ 5'-Phosphate-Dependent Exonuclease (TER51020), which gave similar results