GSM4115895: PV3-M; Mus musculus; RNA-Seq
To isolate interneurons with tdTomato expression, PFC was rapidly dissected placed in digestion buffer (PBS/10% Papain). Following 20 min incubation at 37ºC, digestion was stopped with FACS buffer (PBS/1% BSA), followed by gentle dissociation with a glass Dounce homogenizer (Kontes Glass Company, Vineland, NJ). Single-cell solutions were spun down at 1000 rpm for 5 minutes, re-constituted in 600 ul of FACS buffer, and filtered (40 um) into flow tube. Samples were sorted on a FACS Aria (Yale Immune Monitoring Core Facility) into lysis buffer; average yield was ~150,000 cells/sample. RNA was obtained with the Norgen Single Cell RNA Purification Kit (#51800; Norgen Biotek Corp., Thorold, Ontario, Canada). 100 ng of total mRNA was processed using the Pico RiboGone-Mammalian kit (TakaraBio, Kyoto, Japan) before cDNA synthesis.
GEO Accession
GSM4115895