GSM5182939: scRNA-seq - germ cell AR2481 [HHHNYBBXY_2_IDT-DUI-NXT-318]; Mus musculus; RNA-Seq
Cells were dissociated using 0.05% Trypsin-EDTA (Thermo Fisher Scientific, 25300054) and then for PGCLCs additionally stained with SSEA-1 eFluor 660 (Thermo Fisher Scientific, 50-8813-42) and CD61-PE-Vio770 (Miltenyi Biotec, 130-102-627) for 1h at 4°C. Cells were washed once with 0.5% BSA in PBS and then FACS sorted using a BD FACSAria II SORP or a BD Influx using a 100 um nozzle.  RNA was extracted using phenol-chloroform (Sigma Aldrich, P2069) followed by ethanol precipitation or directly lysed in SMART-seq2 buffer for single-cell sequencing. Bulk total RNA-Seq libraries were prepared using the TruSeq Stranded Total RNA Library Preparation Kit (Illumina, 20020597) followed by paired-end sequencing (2 x 125 bp) on an Illumina HiSeq 2500. Single-cell RNA-seq libraries were prepared using an updated SMART-seq2 protocol with paired-end sequencing (2 x 100 bp) on an Illumina HiSeq 2500. Bulk libraries were sequence in paired-end (2 x 125 nt) on an Illumina HiSeq 2500. Single-cell libraries in paired-end (2 x 100 nt) on an Illumina HiSeq 2500.
GEO
GSM5182939