Bacteriophage titration of E. coli in vivo panel with spotting assay (PSA)
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OVERVIEW
DOCUMENTS/REFERENCES
BIOLOGICAL INFORMATION
BACKGROUND INFORMATION
The phage spotting assay (PSA) for phage titration is used to determine the concentration of a phage product measured as plaque-forming units per mL against a panel of E. coli strains (ATCC 25922, ATCC 700336, AR Bank #0349, and AR Bank #0055).
NUMBER OF ASSAYS ON PANEL
4
ASSAYS ON PANEL
| FAMILY | NAME | ITEM NUMBER | PROTEIN NAME | ORGANISM | ASSAY SUB TYPE |
| Bacteriophage titration of E. coli (ATCC 25922) with spotting assay (PSA) | 697511 | Gram-negative Bacteria | |||
| Bacteriophage titration of E. coli (ATCC 700336) with spotting assay (PSA) | 697512 | Gram-negative Bacteria | |||
| Bacteriophage titration of E. coli (AR Bank #0349) with spotting assay (PSA) | 697513 | Gram-negative Bacteria | |||
| Bacteriophage titration of E. coli (AR Bank #0055) with spotting assay (PSA) | 697514 | Gram-negative Bacteria |
ORGANISM
Gram-negative Bacteria
ASSAY INFORMATION
ASSAY TYPE
Functional
TESTING INFORMATION
PROCEDURE SUMMARY
The testing phage suspension is 10-fold serially diluted with phage dilution buffer (such as SM buffer) for a total of 6 concentrations (typical range is 10⁴ to 10⁹ dilutions). A 0.1 mL aliquot of each phage dilution is mixed with 0.1 mL of bacterial overnight culture in a broth medium. Duplicate phage-bacteria mixtures are prepared with one phage dilution series. Phage absorption to cells is allowed at room temperature for a period of 15 minutes. Molten top agar medium, containing 0.75% agar and with a volume of 3 mL, is added to the phage-bacteria mixture, and the tube is mixed by quick inversion. The contents are plated onto pre-warmed (25°C) bottom agar medium, typically LB containing 1.5% agar. The plates are rotated to evenly distribute the top agar and then allowed to solidify at room temperature. The plates are then incubated for 18 – 24 h at 35-37°C. Plaques are counted on plates with 1 to 300 distinguishable plaques to calculate PFU/mL values.
CRITERIA FOR SIGNIFICANCE
The phage spotting assay (PSA) for phage titration is used to determine the concentration of a phage product measured as plaque forming units per mL (PFU/mL).
TURNAROUND TIME
STANDARD
20 Days
CLINICAL RELEVANCE
THERAPEUTIC AREA
Infectious Disease
ADDITIONAL INFORMATION
BRAND
PDS
TESTING LOCATION
Taiwan - Taipei
ITEM DOES NOT EXIST
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