Triplepure m bio grade

Tissue homogenization was performed using a BeadBug microtube homogenizer (Benchmark Scientific, Edison, NJ, USA). Briefly, a piece of tissue (30–40 mg) was removed from RNAlater and placed in a 2 mL prefilled tube containing 1.5 mm high impact zirconium beads (TriplePure M-Bio Grade; Benchmark Scientific). Lysis buffer (350 µL; RNeasy Mini Kit; Qiagen) was added and the sample was homogenized in the BeadBug for 2 min on the maximum speed. Total RNA was then isolated from the homogenized samples according to the manufacturer’s instructions, eluted with 50 µL RNase-free water, and stored at −80°C until qRT-PCR analyses performed.

The tissue samples were homogenized with a BeadBug homogenizer (Benchmark Scientific, Edison, NJ). Briefly, a piece of ileum and jejunum (30–40 mg) was removed from RNAlater and transferred to prefilled microtubes containing zirconium beads (1.5 mm; TriplePure M-Bio Grade; Benchmark Scientific). Lysis buffer (350 μL; RNeasy Mini Kit; Qiagen) was added, and the tissues homogenized for two minutes at the maximum speed. Each tissue was handled separately. Following homogenization, total RNA was isolated according to the instructions, eluted with 50 μL RNase-free water, and stored at -80°C.