GSM8826320: CTR_Mouse4; Mus musculus; RNA-Seq
Mice were sacrificed and transcardially perfused with PBS. The spleen and the distal colon were collected to perform the total RNA extraction. Cells from the spleen were obtained by smashing on a 70 μm cell (Corning) as long as the sample was completely dissected. The suspension was centrifuged (1000g, 10min, RT) and the pellet lysed in Trizol reagent (Sigma-Aldrich). The distal colon samples (1 cm) were dissected mechanically in Trizol reagent (Sigma-Aldrich) by a cell homogenizer. Isolated total RNA from colon tissue was processed with RNAse-free DNaseI (Invitrogen). Next, total RNA was processed to Nextflex Rapid Directional RNA-Seq kit 2.0 Kit with Nextflex Ribonaut rRNA depletion kit and UDIs (PerkinElmer) . Libraries were prepared with the kit Nextflex Rapid Directional RNA-Seq kit 2.0 Kit with Nextflex Ribonaut rRNA depletion kit and UDIs (PerkinElmer). The libraries were sequenced on Illumina NovaSeq X with 2x150 bp paired-end run; Seq throughput: 50 M PE read per sample; Q30 ≥ 85% .