title: GSM8794085: Tissue, Ighasec+/-, SPF, Ileum, ZE49; Mus musculus; RNA-Seq
library_name: GSM8794085
library_construction_protocol: 1 cm of tissue from the beginning of the duodenum, the middle of the jejunum, the end of the ileum, and the middle of the colon was excised and preserved in RNAprotect (Qiagen) overnight at 4°C. The preserved tissue was then stored at -80°C until processing. Tissue was thawed on ice and transferred to a screw-cap tube containing 450 μL of RLT Plus buffer supplemented with 2-mercaptoethanol and equal quantities of 1.0mm and 0.5 mm zirconium oxide beads (Next Advance). Tissue was homogenized three times for 30 sec at a speed of 6 m/s using Bead Ruptor Elite mill homogenizer (Omni), with 1 min incubation on ice between each cycle. RNA from the lysates was purified using RNeasy Plus Mini kit (Qiagen) with the optional on-column DNase I digest step according to manufacturer protocol. 500 ng of RNA was used as input in the TruSeq Stranded mRNA Library Prep (Illumina) according to manufacturer's specifications.