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- Phage Purification
- March-April
- May-June
- July-August
- September-October
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6.26 Titer
I) Purpose
- Test the concentration of purified T7 phage.
II) Expected Outcome
- Plaques from the bands in the CsCl gradient where phage were purified. Expecting plaques down to at least -5 concentration
III) Reagants Used
- BL21 freezer stock
- LB
- 1.5x top agar
- T7 phage from CsCl gradient
IV) Actual Procedure
- Infect 500 µL of LB with 1 loop-full of BL21 freezer stock and put on shaker at 37◦ C for 5 hours.
- Plate BL21 on an LB plate with 4.5 mL top agar.
- Dilute T7 using a dilution series.
- Spot 2 µL of phage from dilution series onto plate using the concentrations 0, -2, -3, -4, -5, and -6.
- Place in 37◦ C incubator overnight.
V) Results
- The titer of T7 phage showed plaques down to a -5 concentration. This concentration will allow us to perform and EM and examine how pure the phage solution is.
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