title: GSM4200293: Dropseq_LG_Normal_72hAEL_2; Drosophila melanogaster; RNA-Seq
library_construction_protocol: 100 to 150 primary LGs were dissected and kept in 200 µl ice-cooled Schneider's medium (Gibco, 21720024). After dissection, centrifugation at 3,000 rpm and 4 ℃ for 5 minutes was done. Supernatant was discarded and 300 µl of room-temperature Schneider's medium was added to the lymph gland primary lobes. 300 µl of Papain (Worthington, LK003178) pre-heated to 37 ℃, and 4.1 µl of Liberase TM (Roche, 5401119001) were added and gently mixed. The samples were incubated for 20 minutes with gentle nutation. At 5-, 10-, and 15-minute time points of incubation, samples were mixed using 200p pipette. Enzymes were inactivated with 100 µl of ice-cooled Schneider's medium, and samples were kept on ice. Suspended cells were passed through a 40 µm cell strainer (Corning, 352340). After, centrifugation at 3,000 rpm and 4 ℃ for 5 minutes was done. Supernatant was discarded, and 1X filtered sterile PBS was added to cells. Final concentration of cells was fixed to 300 cells/µl for a total of 600 µl. All the Drop-seq and cDNA synthesis methods were followed by previous study (Macosko, Evan Z., et al. "Highly parallel genome-wide expression profiling of individual cells using nanoliter droplets." Cell 161.5 (2015): 1202-1214.). The concentration of beads was fixed to 300 beads/u.  Around 10 minutes Drop-seq run performed for each experiment. single-cell RNA-seq (Drop-seq)
attributes.tag: GEO Accession
attributes.value: GSM4200293