GSM4218162: 5 pooled forager brains, samples collected from Colony O38; Apis mellifera; RNA-Seq
We followed the "short dissociation protocol" from Davie et al. 2018. Briefly, individual bee brains were dissected in ice-cold Dulbecco's phosphate-buffered saline (DPBS, ThermoFisher, cat. #14190144) and cut into small pieces. Pools of five brains were placed in an Eppendorf tube containing 0.05% trypsin-EDTA (Invitrogen, cat. # 25300054) in DPBS. Brains were dissociated on a shaker (25°C, 1000 rpm) for 15 minutes with gentle pipette mixing every 5 min. Debris was removed and cells were spun down at a 400 rcf in a 4°C centrifuge. We removed the supernatant and replaced it with 1 ml of ice-cold DPBS. We repeated this “washing” process twice but, after the third centrifugation, replaced the supernatant with 400 ul of DPBS containing 0.04% UltraPure BSA (ThermoFisher, cat. #AM2616). Cells were then passed through a 40 um Flowmi Tip Strainer (Bel0Art, cat# H136800040). Cell counting was performed on a Nexcelom Cellometer. scRNA-Seq libraries were constructed with the Chromium Single Cell 3' Library and Gel Bead Kit v3 and were sequenced on a 2 x 150nt lane in a NovaSeq6000 (Illumina, San Diego, CA).
GEO Accession
GSM4218162