GSM5849529: germ free; Mus musculus; RNA-Seq
GSM5849529
To isolate single cells, whole epithelial segments or crypts were washed in PBS prior to adding fresh prewarmed Trypsin (Corning, USA) and incubated at 37°C for 50 minutes, with mixing every 20 minutes. Tissues were filtered through a 40 mm mesh cell strainer on ice and pelleted by centrifugation at 4°C and washed twice with ice cold regular medium to remove debris. A manually estimated 4500 single cells per sample were then subjected to 10X Genomics single cell RNA sequencing following manufacturer's recommendations. Illumina HiSeq 3000 was used for deep sequencing. RNA libraries were prepared for sequencing using standard Illumina protocols