The antiviral efficacy
of biopolymer films was determined by following a modified protocol.56 (link) Briefly, each film was placed in individual
wells on a 6-well plate. A total of 10 μL of Phage Phi6 (DSM
21518) stock in Lysogeny broth (LB media and 0.01 M CaCl2) was added to the surface and immobilized across the slide by addition
of a coverslip. This was then incubated for 1 h at room temperature
(20–25 °C). The samples were submerged in 1 mL of LB buffer,
the coverslip was removed, and surfaces were washed with gentle pipetting.
Phi6 submerged in LB was subject to a 10-fold serial dilution. A phage
overlay plate was created by the lower layer LB buffer with 1.5% w/v
Difco Bacterial Agar, overlaid with LB with 0.4% w/v Difco agar, containing
200 μL of Pseudomonas syringae (DSMZ 21482) with a culture optical density of 0.5–0.7. A
total of 10 μL of each dilution was added to the bacterial overlay,
allowed to dry, and incubated for 18 h at room temperature (20–25
°C) prior to the plaque count.
of biopolymer films was determined by following a modified protocol.56 (link) Briefly, each film was placed in individual
wells on a 6-well plate. A total of 10 μL of Phage Phi6 (DSM
21518) stock in Lysogeny broth (LB media and 0.01 M CaCl2) was added to the surface and immobilized across the slide by addition
of a coverslip. This was then incubated for 1 h at room temperature
(20–25 °C). The samples were submerged in 1 mL of LB buffer,
the coverslip was removed, and surfaces were washed with gentle pipetting.
Phi6 submerged in LB was subject to a 10-fold serial dilution. A phage
overlay plate was created by the lower layer LB buffer with 1.5% w/v
Difco Bacterial Agar, overlaid with LB with 0.4% w/v Difco agar, containing
200 μL of Pseudomonas syringae (DSMZ 21482) with a culture optical density of 0.5–0.7. A
total of 10 μL of each dilution was added to the bacterial overlay,
allowed to dry, and incubated for 18 h at room temperature (20–25
°C) prior to the plaque count.