title: Illumina HiSeq 2500 sequencing; 3' RNA-seq of distal colon cells from mice treated or not with DSS and whose mothers were supplemented or not during pregnancy with the prebiotics GOS and inulin
design: 3' RNA-seq of distal colon cells from mice treated or not with DSS and whose mothers were supplemented or not during pregnancy with the prebiotics GOS and inulin
library_construction_protocol: Distal colon fragments were collected then snap-frozen Distal colon samples were lysed in RA1 buffer (Macherey-Nagel) before total RNA extraction with the NucleoSpin RNA kit according to the manufacturer's instructions (Macherey-Nagel). Purified mRNA  were  used for the sequencing The libraries was prepared from 10 ng of total RNA. 15 RNA samples were extracted from hiPS. The mRNA poly(A) tail was tagged with universal adapters, well-specific barcodes and unique molecular identifiers (UMIs) during template-switching reverse transcriptase. Barcoded cDNAs from multiple samples were then pooled, amplified and tagmented using a transposon-fragmentation approach, which enriches for 3'ends of cDNA. 100ng of full-length cDNAs was used as input to the Nextera DNA Sample Prep kit (ref FC-121-1030, Illumina), which enriches for 3′ends of cDNA. Size library is controlled on 2200 Tape Station Sytem (Agilent Technologies).