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5.6 T7+ Liquid Culture Phage Concentration Test
I) Purpose
- To determine if the volume of phage stock added effects the liquid culture concentration result
II) Expected Outcome
- We hope that 100uL of phage will be enough to increase the liquid culture concentration
III) Reagants Used
- E. Coli BL21 overnight from 5.5; T7+ from 5.3 T7 phage amplification/purification; LB from 4.5
IV) Actual Procedure
Liquid culture set up (5.6)
- - Three test tubes were set up. To each, add 4mL of LB and 1mL of E coli BL21 overnight
- - Add 10uL, 100uL, and 1mL to each tube, respectively.
- - Liquid culture was incubated for approximately 48 hours (5.6 4:00pm - 5.8 6:00pm)
Spot test to test amplification results (5.8)
- - Phage liquid culture was purified and spot test was performed using procedures similar to those in 5.3 T7 phage amplification/purification
- - The purified stock was labeled 5.6
- - Spot test plates were incubated overnight
V) Results (5.9)
- - The 10 ul spot test went down to -9, 100 ul to -8, and 1 ml to -7
- - Thus, 10 ul yields the highest titer
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