Team:BYU Provo/Notebook/SmallPhage/Springexp/Period3/Exp/6.27 Mutagen Concentration Test - Third Protocol
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+ | : <u> '''Small Phage''' </u> </font> | ||
: [[Team:BYU Provo/Notebook/SmallPhage/Winterexp|March-April]] | : [[Team:BYU Provo/Notebook/SmallPhage/Winterexp|March-April]] | ||
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: Same process for as Titer to Determine Phage Concentration 1, but plates were incubated for only 24 hours. | : Same process for as Titer to Determine Phage Concentration 1, but plates were incubated for only 24 hours. | ||
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+ | 4) Spot test to confirm phage viability (7.3) | ||
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+ | : Spot test was performed using procedures similar that those in [[Team:BYU Provo/Notebook/SmallPhage/Winterexp/Period3/Exp/4.25 T7 phage viability test|4.25 T7 phage viability test]]] | ||
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+ | : Phage solutions spotted include 2 dilution series from mutagensis result (one created on 6.28 and one newly created from stock). Two positive controls were used: 5.3 stock (this is what we used for mutagenesis) and mutagenized stock from [[Team:BYU Provo/Notebook/SmallPhage/Springexp/Period3/Exp/6.12 Mutagen Concentration Test - Perfected Protocol|6.12 Mutagen Concentration Test - Second Protocol]]. | ||
'''V) Results''' | '''V) Results''' | ||
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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. | : - 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. | ||
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+ | 4) Spot test to confirm phage viability | ||
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+ | : - Both of the two positive controls formed plaques on all of the plates. The only other plaque was on 200ug, new dil, 0. | ||
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+ | : - This confirms our suspicion that phage death occurred because it was incubated for too long with mutagen. | ||
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+ | [[File:BYUSPM3SpotTest.JPG|400px|center]] | ||
'''VI) Conclusion''' | '''VI) Conclusion''' | ||
+ | : Something went seriously wrong with this set of mutagenesis. None of the phage survived. Not even those in the control test tube, with no added mutagen. We weren't able to identify the source of phage death because our phage stock remained viable in the fridge. We have thus decided to repeat this experiment with shorter incubation time to rule out the possibility that it is the longer incubation time that killed the phage. | ||
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Latest revision as of 15:32, 9 September 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)
4) Spot test to confirm phage viability (7.3)
V) Results 1) Applying the mutagen
2) Titer to determine phage concentration
3) Titer to Determine Phage Concentration 2
4) Spot test to confirm phage viability
VI) Conclusion
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