title: GSM3034758: wild-type 1D oligodendrocyte differentiation; Mus musculus; RNA-Seq
library_construction_protocol: Drop-Seq was performed as previously described (Macosko et al., 2015). In brief, three individual syringes containing cell suspension at 200,000 cells/mL) barcoded mRNA capture beads (MACOSKO-2011-10, ChemGenes) in lysis buffer (prepared according to instructions in Drop-Seq protocol (Macosko et al., 2015) at 200,000 beads/mL, and QX200 droplet generation oil (186-4006, Bio-Rad), respectively, were placed in a Legato™ 100 pump (788100, Analytical West). Each output was connected to a custom-made PDMS co-flow microfluidic droplet generation device (Nanoshift) under microscope supervision. Cell and bead flow was set to 4,000uL/hr, and oil flow was set to 15,000uL/hr during droplet generation. These settings were optimized to allow at most one bead per one cell. Droplets were collected and then broken to release beads. First strand cDNA synthesis with incorporation of a PCR handle was performed on the collected beads using the Maxima H Minus Reverse Transcriptase (EP0752, Thermo Fisher). The resulting full-length cDNA library was prepared for sequencing as described (Macosko et al., 2015)
attributes.tag: GEO Accession
attributes.value: GSM3034758