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5.9 T7+ Liquid Culture Phage Concentration Test #2
I) Purpose
- To determine if the volume of phage stock added effects the liquid culture concentration result
II) Expected Outcome
- We expect the same results as from 5.3. We also hope to see 1 ul produce a titer to -10.
III) Reagants Used
- E. Coli BL21 overnight from 5.8; T7+ from 5.6 10 ul stock; LB from 4.5
IV) Actual Procedure
1. Liquid culture set up (5.9)
- Three test tubes were set up. To each, add 4mL of LB and 1mL of E coli BL21 overnight
- Add 1uL, 10uL, and 100uL phage to each tube, respectively.
- Liquid culture was incubated for approximately 48 hours
2. Spot test to test amplification results
- Phage liquid culture was purified using procedures similar to those in 5.3 T7 phage amplification/purification. The resulted purified stock was labeled 5.13 (5.13)
- Spot test was performed using procedures similar to those in 5.3 T7 phage amplification/purification (5.15)
- Spot test plates were incubated overnight
V) Results
- Complete clearage of liquid culture was observed for once
- All spot tests showed plaques to -8, but there was also a lot of contamination -> from the LB plates; the plates have been completely disposed of.
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