Sample 150
Mus musculus
Protocols: Primary fibroblasts were derived from adult CAST/EiJ x C57/Bl6 mice. Tail explants were cultured in fibroblast medium (DMEM high glucose, 10% ES cell FBS, 1% Penicillin/Streptomycin, 1% non-essential amino acids, 1% sodium-puryvate, 0.1mM b-Mercaptoethanol). Cells were passaged twice before distributed as individual cells into wells of 384 or 96 well plates using a FACS. Wells was pre-loaded with lysis buffer. ES cells were cultured as in Meissner et al. Methods Mol Biol 2009, and similarly distributed via FACS into as single cells into wells of 96 and 384-well plates Cell lysis. Smart-seq2, indexed by Nextera i5 and i7 and with a unique molecular identifier present in template-switching oligonucleotide. No exogenous spike-ins were used..