All solutions were degassed by purging with N2 and with rigorous stirring for a minimum of 2−3 h (1 L of buffer in a plastic flask with minimum head space) before placing in an anaerobic cabinet (Belle Technology; ≤2 ppm O2) overnight at ca. 25 °C.
Next to the bike lane, which will be 10 feet wide, will be an eight-foot section of pavement that will act as a buffer, with plastic posts and large planters to keep cars from entering.
In brief, flies were mechanically homogenized with a plastic pestle in homogenization buffer (100 mM Tris-HCl [pH 8.8], 0.5 mM EDTA, 1.0% SDS).
Thoracic muscle (~1-2 mg) was homogenized with a plastic pestle in 1 ml of buffer containing 0.25 m sucrose, 10 mm EDTA, and 30 mm Tris (pH 7.6) and centrifuged at 1000 g for 5 min at 4°.
Homogenates of juvenile trout were prepared by adding homogenization buffer (4 times the sample weight; PBS + 2 TIU aprotinin, C. Roth, Germany), mixing with a plastic pestle, centrifuging (10 min, 4 °C, 20,000×g Eppendorf 5810R) [17] and storing the supernatants at −80 °C.
For DLS, reactions of SbpA in self-assembly buffer (100 μL) were pipetted into a disposable solvent resistant microcuvette (Malvern Instruments, Worcestershire, United Kingdom), sealed with a plastic lid and PARAFILM, and then placed into the instrument.
The buffer was withdrawn and replaced with 2 ml HMS, and the cells were scraped from the flask surface with a plastic cell scraper.
In brief, the tissue was homogenized with a plastic tissue grinder and pestle (Kontes Glass Company, Vineland, NJ, USA) in 350 µl lysis buffer.
The precipitated DNA strands were removed with a plastic spatula or pipette and transferred to a 1.5 mL microcentrifuge tube containing 100 200 pl TE buffer (10 mM Tris-HCl and 0.2 mM Na2EDTA, pH 7.5).
CT scans of rabbit's cranium soaked in 10% buffered formalin inside a plastic recipient can be made with the head positioned in a natural way, hugged by wax and supported by wood to keep the acquisition stable [ 11].
