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- Small Phage
- March-April
- May-June
- July-August
- September-October
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5.5 Amplification from a plaque test
I) Purpose
- Test the procedure for creating phage liquid culture from a single plaque on a plate
II) Expected Outcome
- In spot test, we should see plaques until the phage dilution series became too diluted.
III) Reagants Used
- E. Coli BL21 overnight from 5.4; ×6 plate from 5.3 T7 phage selection method test; LB from 4.3
IV) Actual Procedure
1. Amplifying phage from a single plaque (5.5)
- To a test tube, add 1mL of E coli BL21 overnight (5.4) and 4mL of LB
- Stab one of the plaques on a ×6 plate from the 5.3 T7 phage selection method test with a 1mL pipet tip, puncturing the agar. The plaque in question is labeled with a circle.
- The pipet tip is then dipped into the overnight/LB solution. The solution is then pipetted up and down several times to maximize the amount of phage that gets into the solution.
- The solution was then incubated for approximately 24 hours (5.5 noon - 5.6 2:00pm )
2. Spot test to test amplification results (5.6)
- Phage liquid culture was purified and spot test was performed using procedures similar to those in 5.3 T7 phage amplification/purification. Stock created was labeled 5.4
- Spot test plates were incubated overnight (5.6 4:00pm - 5.7 12:00pm )
V) Results (5.7)
- Our method of amplification from a single plaque was successful. Phage plaque was observed up to -7
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