PROTOCOL
The best way to Isolate bacteriophage DNA
As someone who has conducted bacteriophage research, I understand how common it is to encounter host DNA contamination when isolating a phage…
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Plaque-Based Optimization of the Phages Lytic Characteristics
The isolates of wild lytic phages from the transient stocks are propagated with the corresponding host bacterial isolates and the representative standard…
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Production of the Transient Phage Stock
Procedure Cut 5–10 plaques with a sterile Pasteur pipette and place them in 1.5 ml sterile Eppendorf micro-centrifuge tubes.Add into 300 µl of…
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Bacterial Stock Culture preparation
ProcedurePrior to the procedure, a stock culture of the phage target bacteria should be maintained on any general media preferably Trypticasein soy…
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Preparation of anti-phage agent
ProcedureImmediately prior to use, transfer 3.3 ml of 13% PRE added to 7 ml of freshly prepared ferrous sulphate solution (0.01%). After…
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Preparation of 13% Pomegranate Rind Extract (PRE)
ProcedureBlend pomegranate rind in distilled water (25% w/v) and boil for 10 min.Centrifuge (20,000xg, 4 °C, 30 min) (A further purification of…
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Preparation of FeSO4.7H2O Solution
ProcedureFirst freshly prepare a solution (0.01%) of FeSO4.7H2O in Lambda buffer, pH 6.3.Sterilized by membrane filtration (0.45 lm, Whatman)
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Preparation of SM buffer (lambda diluent)
SM buffer is primarily used in biology laboratories, and its main function, like any other buffer, is to stabilize the pH. That…
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Preparation of Phosphate Buffered Saline (PBS)
Phosphate-buffered saline (abbreviated PBS) is a buffer solution commonly used in biological research including phage work. It is a water-based salt solution…
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Common media used in phage work
microbiological MediaList of the media usedLuria broth (LB): tryptone 10 g l−1 (HiMedia, Mumbai, India), yeast extract5 g l−1 (HiMedia, Mumbai, India),…