GSM3294752: P1-5_RA-Replicate2; Mus musculus; RNA-Seq
Testes from postnatal day 1.5 (P1.5) mice (12hr after treatment at midnight on P1) were used to generate suspensions of cells following enzymatic digestion as described previously (Ogawa et al., 1997; Nagano et al., 2003; Oatley and Brinster, 2006). Briefly, testicular parenchyma from at least two pups were pooled and digested with 0.25% trypsin/EDTA (ThermoFisher Scientific) for ~6-8 minutes at 37oC, quenched with 10% FBS (v/v, Sigma). Testis cell suspensions were used directly for single-cell RNA-seq facilitated by the 10X Genomics Chromium Drop-Seq approach. Briefly, Cell suspensions were loaded into Chromium microfluidic chips with 3' v2 chemistry and used to generate single-cell gelbead emulsions (GEMs) with using the Chromium controller (10X Genomics) per manufacturer recommendations (Zheng et al., 2017). In all cases, suspensions containing ~17400 cells were loaded on the instrument with the expectation of collecting up to 10,000 GEMs containing single cells. Single-cell 3' libraries were prepared according to manufacturer recommendations (10X Genomics v2 chemistry).
GEO Accession
GSM3294752