Team:BYU Provo/Notebook/SmallPhage/Springexp/Period2/Exp/5.20 Mutagen Concentration Experiment
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: - 6uL of 5.3 phage stock was added to each centrifuge tube. ''From 5.15 Titer Test on 5.3 T7 new Phage Stock, we know that 5.3 phage stock has 7E8 particle/20uL. Thus, to each of the test tubes, we are adding approximately 2.1E8 particle. This gives us a multiplicity of infection lower than 0.5 particles per cell.'' | : - 6uL of 5.3 phage stock was added to each centrifuge tube. ''From 5.15 Titer Test on 5.3 T7 new Phage Stock, we know that 5.3 phage stock has 7E8 particle/20uL. Thus, to each of the test tubes, we are adding approximately 2.1E8 particle. This gives us a multiplicity of infection lower than 0.5 particles per cell.'' | ||
- | : - Incubate phage on shaker at 37C for approximately | + | : - Incubate phage on shaker at 37C for approximately 30min. |
: - Liquid culture from the five test tubes is then centrifuged at 4000 rpm for 10 minutes to pellet cell debris. Supernatant containing phage particles is removed and 1mL of chloroform is added destroy any remaining E coli cells. | : - Liquid culture from the five test tubes is then centrifuged at 4000 rpm for 10 minutes to pellet cell debris. Supernatant containing phage particles is removed and 1mL of chloroform is added destroy any remaining E coli cells. | ||
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: We plated -4 dilution titers with x6 and x8 top agar using similar procedure to the previous selection test. | : We plated -4 dilution titers with x6 and x8 top agar using similar procedure to the previous selection test. | ||
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+ | 6) plating 45 plates + plaque test | ||
'''V) Results''' | '''V) Results''' |
Revision as of 21:00, 12 June 2013
Small Phage May - June Notebook: Experiments
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5.20 Mutagen Concentration Experiment
I) Purpose
II) Expected Outcome
III) Reagents Used
IV) Procedure 1) Applying the mutagen (5.20)
2) Spot test (5.22)
3) Preliminary selection test (5.24)
4) Preliminary selection test - second round (5.29)
5) Preliminary selection test - third round (6.3)
6) plating 45 plates + plaque test V) Results 1) Applying mutagen
2) Spot test
3) Preliminary selection test
4) Preliminary selection test 2
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