GSM4059128: GF_3 [RNA-seq]; Mus musculus; RNA-Seq
For epithelial cell preparation from colon, colon was flushed with ice-cold PBS, cut open longitudinally. Distal and proximal colon region were processed separately. Each part was minced into small pieces in cold PBS. Enterocytes were mechanically isolated by shaking in PBS with 30 mM EDTA, at 37oC. Crypts were recovered and then stored at -80°C for RNA extraction. Only samples from distal colon were used in our study. Total RNA from the samples was isolated using the Fibrous Tissue Kit (Qiagen) according to the manufacturer's instructions. The Poly(A)Purist™ MAG Kit (Ambion) was used for mRNA selection. Sequencing libraries were prepared using 1µg of DNAse-treated total RNA in the first step of the TruSeq RNA Sample Preparation v2 protocol (Illumina, Part# 15026495 Rev. A) as recommended by the manufacturer.
GEO Accession
GSM4059128