Large-scale amplification of and preparation coated surfaces
phage was grown and purified following standard biochemical protocol. An amount of 500 mL E. coli TG1 culture was grown in 2ytx media to mid-log phase and infected with 1 mL wild-type bacteriophage (1012 PFU/ mL). The culture was incubated at 37°C with shaking for five to six hours and centrifuged at 8000g for 30 minutes to remove bacterial cells; and then, the virus was collected by subsequent centrifuging at 20000 g for 150 minutes. The resultant pellet was suspended in 500 μL PBS and concentration of the isolated bacteriophage was determined spectrophotometrically using an extinction coefficient of 3.84 cm2/mg at 269 nm.
Preparation of samples for TEM analysis
A solution of isolated M13 phage (20 μL) was adsorbed into a 300-mesh carbon-coated copper grid (AGS160-3) for two minutes. For transmission electron microscopy (TEM) analysis, the grid was then stained with 2% uranyl acetate. A Zeiss - EM10C - 80 KV (Oberkochen, Germany) transmission electron microscope was employed for visualizing the prepared samples.