title: GSM8072289: Colon, Il10KO W8, Immune fraction, gene expression; Mus musculus; RNA-Seq
library_construction_protocol: Tissue biopsies (approximately 0.5 cm long) were collected from the distal colon and stored at -80°C in RNAlater (Invitrogen). To extract RNA, tissue was lysed in RLT Plus Buffer (QIAGEN) + 2% b-mercaptoethanol by bead-beating (Precellys). RNA was isolated with the RNeasy Plus Mini Kit (QIAGEN) according to the manufacturer's instructions and quantified using a Nanodrop spectrophotometer (ThermoFisher). Libraries were constructed following the manufacturer's guidelines (Chromium Next GEM Single Cell 3ʹ v3.1 (Dual Index) with Feature Barcode technology for Cell Surface Protein (10X Genomics)), with TotalSeq-derived barcodes being retained only for the immune cell fractions. Sequencing was performed by Novogene Co. Ltd (UK) on NovaSeq 6000 sequencers with a targeted sequencing depth of 300M reads per gene expression sample and 50M reads per TotalSeq-derived sample. Read length was PE150 (Novogene) or 28-10-10-150 for gene expression samples and 28-10-10-25 (NGI) for TotalSeq-derived samples.