RNAseq_PosteriorGut_Winter_10weeks_Control6
RNA was extracted from 60 mg of spleen or posterior intestine tissue. Tissues were homogenized in 0.5ml of TRI Reagent solution (Invitrogen) using Lysing Matrix D tubes (MP Biomedicals). Homogenization was performed in a FastPrep homogenizer (MP Biomedicals) using 1 cycle of 30 s at 6 m/s. Then, total RNA was isolated using the MagMAX-96 for Microarrays Total RNA Isolation Kit (Applied Biosystems) according to the manufacturers instructions. RNA concentration and purity were measured using a Nanodrop 2000c (Thermo Scientific). Finally, RNA quality and integrity were measured on an Agilent Bioanalyzer 2100 total RNA Nano series II chip (Agilent). RNA integrity number (RIN) values were between 8 and 10. Sequencing Illumina libraries were prepared following standard protocols in GeneWiz (Azenta Life Sciences) and sequenced in NovaSeq in a 2x150 configuration.