RNA-Seq of Mus musculus: Colon organoids IL-22-3
3' Adenylation of DNA fragments. Sequencing adapter ligation (utilizing T-A pairing of adapter and DNA fragments). Amplification of library via PCR (13 cycles) (amplification of "unmarked" 1st strand only). Libraries are assessed using either Agilents Bioanalyser DNA 1000 chip or TapeStation D1K TapeScreen system (please refer to the attached PDF for a summary of the library sizes). We use qPCR to quantify individual libraries before normalizing (2nM) and pooling. Libraries were pooled and clustered through the Illumina cBot system using TruSeq PE Cluster Kit v3 reagents followed by sequencing on the Illumina HiSeq 2500 system with TruSeq SBS Kit v3 reagents with 117 (101 read 1, 16 cycles index read).