GSM8878031: PBS-IV-Replicate-3; Mus musculus; RNA-Seq
RNA from flash frozen colonic tissues was extracted using RNeasy Plus Universal mini kit (Qiagen). The integrity of the isolated RNA was analyzed using the Agilent TS HS RNA Kit and TapeStation 4200 at the Genome Technology Center at the Azrieli Faculty of Medicine, Bar-Ilan University. 1000 ng of total RNA was treated with the NEBNext poly (A) mRNA Magnetic Isolation Module (NEB, #E7490L). RNA sequencing libraries were produced by using the NEBNext Ultra II RNA Library Prep Kit for Illumina (NEB #E7770L).  Quantification of the library was performed using a dsDNA HS Assay Kit and Qubit (Molecular Probes, Life Technologies) and qualification was done using the Agilent TS D1000 kit and TapeStation 4200. 250nM of each library was pooled together and diluted to 4nM according to the NextSeq manufacturer's instructions. 1.6pM was loaded onto the Flow Cell with 1% PhiX library control. Libraries were sequenced with the Illumina NextSeq 550 platform with single-end reads of 75 cycles according to the manufacturer's instructions.