After this, 1 mmol of maghemite nanoparticles (γ-Fe2O3) were mixed with the BSA containing buffer and the mixture was vortexed for 1 h at room temperature.
Pellets were macerated in 1 ml buffer and vortexed, then the entire suspension was plated on Potato Dextrose Agar (PDA; 9.5 cm diameter Petri dish) supplemented with the antibacterial chloramphenicol.
The suspension was washed three times with PBS, resuspended in Laemmli buffer and the bound proteins were eluted by vortexing during 10 min. Ku70, Ku80 and AP-2 were revealed by western blotting.
Briefly, after evaporation of the eluent the fractions were reconstituted in 50μL PBS buffer and vortex mixed for 1min.
Aspirate the supernatant and add 1 ml cold RIPA or IPP150 buffer and vortex for 30 seconds.
For clinical isolates tests, fast DNA extraction was performed using glass beads (in Tris-EDTA buffer) and vortex agitation [26].
The remaining material, scraped into 0.5 ml MES buffer and vortexed for 1 min, constituted the insoluble fraction.
The pellets were eluted in protein loading buffer, and samples were vortexed and boiled for 5 mins at 95°C.
The pellet was washed once in hypotonic buffer and resuspended by vortexing in hypertonic buffer (20 mM HEPES pH 7.9, 420 mM NaCl, 1.5 mM MgCl2 0.1 mM EGTA, protease inhibitors as per hypotonic buffer above).
The supernatant was removed, the pellet was dissolved in 1ml of CE buffer, and samples were vortexed for about 15 sec.
