title: GSM5792068: AB9215; Mus musculus; RNA-Seq
library_construction_protocol: Kidney single cell suspension: renal capsule was removed and the tissue minced with razor blades into 1 mm pieces. Each minced kidney was placed in 1 ml DMEM medium containing collagenase IV (Worthington Biochemical Cat no. LS004188, final concentration 0.25 mg/ml), neutral protease (Worthington Biochemical Cat no. LS02106, final concentration 0.25 mg/ml), and DNase (Roche Cat. no. 10104159001, final concentration 2000 U/ml), and incubated in a thermomixer at 37°C, 300 RPM, for 25-30 min. During this time, the suspension was passed 10 times through a 18G syringe and 8 times a 21 G syringe. Single-cell RNA-seq libraries were prepared as previously described {Jaitin, 2014}. In brief, mRNA from single cells sorted into capture plates were barcoded and converted into cDNA and then pooled using an automated pipeline. The pooled sample was linearly amplified by T7 in vitro transcription, and the resulting RNA was fragmented and converted into a sequencing-ready library by tagging the samples with pool barcodes and Illumina sequences during ligation, RT, and PCR. Each pool of cells was tested for library quality and concentration as described previously {Jaitin, 2014}