title: GSM7830922: BLA, p56_1; Mus musculus; RNA-Seq
library_construction_protocol: Mice were deeply anesthetized by i.p. injections of 200 mg/kg Ketamine and 40 mg/kg Xylazine, and perfused with 10 mL ice-cold Sucrose-HEPES Cutting Buffer containing (in mM) 110 NaCl, 2.5 KCl, 10 HEPES, 7.5 MgCl2, and 25 glucose, 75 sucrose (~350 mOsm/kg), pH=7.4 2.All the solutions/reagents were kept on ice in the following procedures unless otherwise specified. The brain was extracted and cut (300 μm) on a vibratome (Leica VT1000S, Germany) in cutting buffer, and the slices were transferred into a  Dissociation Buffer  containing (in mM): 82 Na2SO4, 30 K2SO4, 10 HEPES, 10 glucose and 5 MgCl2, pH=7.4 2.BLA was microdissected under a microscope (Olympus SZX10) covering the anterior (Bregma −0.59) and posterior (Bregma −3.0) for adult BLA and the anterior (Bregma −0.8) and posterior (Bregma −2.8) for juvenile BLA. The protocol for single nucleus isolation was optimized from previous studies (Matson KJE, et al. Isolation of Adult Spinal Cord Nuclei for Massively Parallel Single-nucleus RNA Sequencing. J Vis Exp (2018). Nanoliterscale Gel Beads-in-emulsion (GEMs) generation, barcoding, cDNA amplification, and library preparation were done using the Chromium Next GEM Single Cell 3' Reagent Kits v3.1 according to the manufacturer's protocol.