After gentle homogenization with a Dounce homogenizer, cell lysates were subjected to differential centrifugation.
All samples were extracted twice with equal volumes chloroform (1 min homogenization with the PRO200 Homogenizer and 30 min centrifugation at 10,500 × g).
Water at neutral pH is added to the tissue (1.0 mg/100.0 mg of tissue), followed by thorough homogenization with a tissue homogenizer.
After homogenization with a dounce homogenizer B pestle, samples were centrifuged twice (2500 g, 5 min, 4°C) to remove nuclei and centrifuged again (10 000 × g, 10 min, 4°C) to obtain the heavy membrane pellet.
Frozen hepatic tissue was disrupted by homogenization with a rotor stator homogenizer, and total RNA was isolated with Qiagen RNeasy columns Qiagen Inc., Valencia, CA).
Cells were lysed using a nitrogen bomb (Parr) at 200 psi for 10 min, followed by mechanical homogenization with a glass glass dounce homogenizer.
Brain homogenates from each animal were prepared as whole homogenates of brainstem sections by bead homogenization with 1.0 mm silica beads in 1X Dulbecco's PBS pH 7.4 (lacking Ca2+ and Mg2+).
After incubation on ice and homogenization with 45 strokes with a Dounce homogenizer, the extracts were centrifuged for 30 min at 4°C.
After simple plant leaf and callus tissue homogenization with DNA extraction buffer using a hand-operated homogenizer, the leaf and callus cells were lysed with 20% SDS.
Cell nuclei were resuspended in buffer A, centrifuged (1000× g, 10 min, 4 °C) and opened by gentle homogenization with three volumes of buffer A in Dounce homogenizer.
Two additional homogenization with MeCN and filtration were performed.
