GSM5901258: Wildtype colon - mouse 4; Mus musculus; RNA-Seq
Colons were dissected, manually stripped of myenteric layers, and enzymatically digested in Liberase TM/DNAse I. Dissociated tissue was strained through 70 and 40-micron filters, and the resulting single-cell suspension was treated with a red blood cell lysis buffer. Dead cells were removed using a magnetic separation kit with antibodies targeted against annexin V. The purified cellular suspension was submitted for microfluidic processing. All samples were processed on the same day, representing a single batch run. Cells were washed with PBS and pre-loading viability was confirmed to exceed 75%. Cells were run on the Chromium (10X Genomics) platform for single-cell lysis, reverse transcription, barcoding, and library preparation. Paired-end sequencing of 3' transcript ends was performed on a HiSeq machine (Illumina).