GSM8603354: Adult-CD8-4; Mus musculus; miRNA-Seq
Spleens from adult or neonatal mice were collected in RPMI+10% FBS and mashed through a 40µM cell strainer to create a single-cell suspension. Spleens underwent magnetic enrichment with CD8a and CD4 MicroBeads (Miltenyi Biotec; cat# 130-117-044; cat# 130-117-043), respectively, per the manufacturer's instructions. Total RNA was isolated with Trizol, with an extra chloroform extraction to remove residual phenol and glycoBlue as a carrier to improve recovery. Libraries were prepared with NEBNext Small RNA Library Prep Kit (New England Biolabs) using 20-100ng total RNA as input, size selected for insert sizes ~18-35bp. small RNA-seq