Gelatin agarose

Growth medium for bacterial cultures (LB broth, LB agar, EMB agar, SS agar, TCBS agar, Deca strength phage broth, Mueller Hinton Agar) and Bovine Serum Albumin (BSA) were purchased from Himedia Laboratories. DMEM medium, agarose resins (Gelatin–agarose, Heparin agarose and Sepharose 4B), Resazurin and Triton X 100 were procured from Sigma Aldrich. Fetal Bovine Serum (FBS), Penicillin-Streptomycin mixture (PenStrep from Gibco), Amphotericin B and SYBR green master mix 2X DyNAmo Color Flash were obtained from Gibco, Thermo Fisher Scientific. All other salts and other reagents were procured from Sigma Aldrich.

Gelatin-agarose (≥3 mg/ml of agarose beads) and Heparin agarose beads (54-210 USP/ml of agarose beads) from Sigma Aldrich were used to study the binding of bacteriophages to gelatin and heparin. 100 μl of agarose beads and Heparin/Gelatin coated agarose beads were mixed with 200 μl of phage lysate (titer 10⁶) and incubated for 1 h under shaking condition at 37 °C. The resins were allowed to settle down and excess lysate was removed. The resins were washed three times with 1 ml PBS to remove the non-adherent phages. The resins were then resuspended in equal volumes of E. coli culture and the absorbance at 600 nm was measured at different time intervals.