Zahrah Adnan Al-shmmary
hi when isolate phage you must centrifuge the sample which contain phage twice then added to bacteria staphylococcus and put at 37 c .you can use buffer in pH 7
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John Harvey Santos
Hi Assefa Asnakew Abebe ! To isolate lytic phages against Staphylococcus spp., there are a few key factors you need to consider:
- Optimal pH: The pH range of 6.5 to 7.5 is generally suitable for the growth of Staphylococcus spp. and phage isolation. Adjusting the pH within this range is recommended.
- Concentrations of MgSO4 and CaCl2: The typical concentrations of MgSO4 and CaCl2 in phage isolation media can vary, but a commonly used recipe is 10 mM MgSO4 and 10 mM CaCl2. These divalent cations can aid in phage adsorption and stability.
- Soft agar amounts: The amount of soft agar in the isolation process depends on the specific protocol being used. Generally, a concentration of 0.4% to 0.7% agar is employed for phage plaque formation. This concentration allows for the diffusion of phages while maintaining their localized growth.
- Sample sources: For phage isolation against Staphylococcus spp., potential sample sources can include environmental samples (e.g., soil, wastewater), clinical samples (e.g., wound swabs, nasal swabs), or even existing phage collections. These sources may contain phages that infect Staphylococcus spp.
- Gelatin in SM buffer: Gelatin is commonly included in SM (saline-magnesium) buffer used for phage storage and propagation. It acts as a stabilizer, protecting phage particles from degradation by nucleases. However, its use is not mandatory, and alternative stabilizers, such as BSA (bovine serum albumin), can also be employed.
Hope these info helped you! It will be great to check published literatures, too.
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