Team:BYU Provo/Notebook/SmallPhage/Springexp/Period3/Exp/6.27 Mutagen Concentration Test - Third Protocol
From 2013.igem.org
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: - The stock solutions are now stored in 4 Celsius. | : - The stock solutions are now stored in 4 Celsius. | ||
- | 2) Titer to | + | 2) Titer to Determine Phage Concentration 1 (6.28) |
: - For each mutagenesis, specifically 0, 100, 200, and 500ug/mL, 1:100 dilution series were performed to generate 0, -2, -4, -6, and -8 dilutions. | : - For each mutagenesis, specifically 0, 100, 200, and 500ug/mL, 1:100 dilution series were performed to generate 0, -2, -4, -6, and -8 dilutions. | ||
- | : - For each dilution, a test tube was assigned to it. To each test tube, 0.75mL of BL21 liquid culture overnight was added and then transfected with 30uL of phage dilution. After 20 minutes of incubation, 7mL of x8 agar was added to each test tube and the content was then plated | + | : - For each dilution, a test tube was assigned to it. To each test tube, 0.75mL of BL21 liquid culture overnight was added and then transfected with 30uL of phage dilution. After 20 minutes of incubation, 7mL of x8 agar was added to each test tube and the content was then plated. |
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+ | : Plates were incubated upside down for 72 hours. | ||
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+ | 3) Titer to Determine Phage Concentration 2 (7.2) | ||
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+ | : Same process for as Titer to Determine Phage Concentration 1, but plates were incubated for only 24 hours. | ||
'''V) Results''' | '''V) Results''' | ||
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2) Titer to determine phage concentration | 2) Titer to determine phage concentration | ||
+ | : - The plates were left in for too long so the bacteria overtook the plaques that formed; no viable data. | ||
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+ | 3) Titer to Determine Phage Concentration 2 | ||
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+ | : - There was one plaque on the "500ug -2" plate and 11 plaques on the "200ug -8" plate. All other plates had no plaques. We are not sure why this is. | ||
'''VI) Conclusion''' | '''VI) Conclusion''' |
Revision as of 20:22, 3 July 2013
Small Phage May - June Notebook: Experiments
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6.27 Mutagen Concentration Test - Third Protocol
I) Purpose
II) Expected Outcome
III) Reagents Used
IV) Procedure 1) Applying the mutagen (6.12)
2) Titer to Determine Phage Concentration 1 (6.28)
3) Titer to Determine Phage Concentration 2 (7.2)
V) Results 1) Applying the mutagen
2) Titer to determine phage concentration
3) Titer to Determine Phage Concentration 2
VI) Conclusion
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