Difference between revisions of "M13 Bacteriophage Production Protocol"
From GcatWiki
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| + | == Preparing Phage == | ||
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1. Grow 3mL of cells with phage plasmid in liquid media overnight. These cells will produce M13 phage which can then be isolated. For example, grow J100265 + J100270 S1030 cells in 50ug/mLCarb + 50ug/mL Kan + 10ug/mL Tet for SPT7-spec phage. | 1. Grow 3mL of cells with phage plasmid in liquid media overnight. These cells will produce M13 phage which can then be isolated. For example, grow J100265 + J100270 S1030 cells in 50ug/mLCarb + 50ug/mL Kan + 10ug/mL Tet for SPT7-spec phage. | ||
2. Centrifuge 1.5mL of cells at full speed (~15,500 RPM) for at least two minutes. The cells will be spun down into a pellet, while the phage will remain in the supernatant. Save the supernatant in a separate microfuge tube. The phage will remain viable if stored at 4C. | 2. Centrifuge 1.5mL of cells at full speed (~15,500 RPM) for at least two minutes. The cells will be spun down into a pellet, while the phage will remain in the supernatant. Save the supernatant in a separate microfuge tube. The phage will remain viable if stored at 4C. | ||
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| − | Host | + | == Preparing Top and Bottom Agar == |
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| + | == Plating Top Agar with Phage and Host Cells == | ||
Revision as of 21:54, 17 May 2017
Preparing Phage
1. Grow 3mL of cells with phage plasmid in liquid media overnight. These cells will produce M13 phage which can then be isolated. For example, grow J100265 + J100270 S1030 cells in 50ug/mLCarb + 50ug/mL Kan + 10ug/mL Tet for SPT7-spec phage.
2. Centrifuge 1.5mL of cells at full speed (~15,500 RPM) for at least two minutes. The cells will be spun down into a pellet, while the phage will remain in the supernatant. Save the supernatant in a separate microfuge tube. The phage will remain viable if stored at 4C.
