Beadbeating was increased to 2 × 1.5 min at 50 Hz using a tissue lyser (LT, Qiagen, Hilden, Germany) DNA yield was assessed using Qubit fluorometric analysis (Thermo Fisher Scientific, Waltham, MA, USA).
Tissues were homogenized in PBS using a Tissue Lyser homogenizer (Quiagen).
DRG were lysed in RLT® buffer using a tissue lyser (Qiagen Doncaster, Australia), RNA was extracted (RNeasy Mini-kit, Qiagen) and the quantity and quality of the RNA determined (Nanodrop, Thermo Scientific, USA, Bioanalyzer, Agilent, USA).
Stool specimens were disrupted and homogenized in buffer ASL of the Stool kit with 5 mm stainless steel beads using a Tissue lyser (Qiagen) at a rate of 25 times/sec for 2 min, followed by incubation at 95°C for 5 min. The Stool kit was also used for DNA extraction from the biopsy specimens.
Left and right kidney from each animal were homogenized using a Tissue Lyser (Qiagen, TissueLyser Type mm 301), UV radiated 2 metal beats (3 mm), and 30 HZ in 2 min. 100 µl of 10 fold dilutions (range undiluted – 1∶1000) of the tissue homogenate was used to inoculate Sabouraud agar and colonies were counted at day 2 for CFU determination.
For the determination of virus titers on the indicated days post-infection, the ovaries were removed and homogenized in medium using a Tissue Lyser (Qiagen), and the spleens were made into a single cell suspension between two frosted slides and resuspended in 10 ml complete RPMI-1640 medium.
Approximately 40 mg of tissue was homogenized using a Tissue-lyser (Qiagen) in ice cold RIPA buffer (with protease inhibitor cocktail) and centrifuged at 13,000 g for 15 min at 4°C and the pellet discarded.
Human muscle samples were homogenized (n = 13) using a Tissue-lyser (Qiagen, Crawley West Sussex, UK) in 50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1 mM EGTA, 1 mM EDTA, 0.25% NaDeoxycholate, 1% Triton X-100.
Samples of ∼100 mg dry tissue were placed in 2 ml Eppendorf vials and homogenized in 1 ml 50 mM phosphate buffer (pH 7.4) using a Tissue-lyser II with a steel bead at 30 Hz for 20 seconds (Qiagen, Copenhagen, Denmark).
For RNA extraction from total muscles, freshly harvested tissue was disrupted using a Qiagen Tissue Lyser (Qiagen, Basel, Switzerland) in 1 ml TRIzol Reagent for 100 mg of tissue (Life Technologies).
