Team:BYU Provo/Notebook/SmallPhage/Springexp/Period3/Exp/7.17 Mutagen Concentration Test - Fifth Protocol
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2) Spot Test to Confirm Phage Viability (7.19) | 2) Spot Test to Confirm Phage Viability (7.19) | ||
- | - Add 0.75mL of BL21 overnight into 6 test tubes | + | : - Add 0.75mL of BL21 overnight into 6 test tubes |
- | - Add 7mL of x1 top agar to each tube and plate it on top of the 6 LB plates | + | : - Add 7mL of x1 top agar to each tube and plate it on top of the 6 LB plates |
- | - Divide the plates into 6 quadrants with a sharpie. | + | : - Divide the plates into 6 quadrants with a sharpie. Label the quadrants 7.15, -4, -5, -6, -7, and -8. |
- | - Using the mutagenized phage stock from step 1 above, create a 1: | + | : - Using the mutagenized phage stock from step 1 above, create a 1:100 dilution series of the 6 solutions in step 1 for -2 and -4 in eppendorf tubes. Then make a 1:10 series for -5, -6, -7, and -8. |
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+ | : - Spot 5ul of each eppendorf tube (-4 through -8) onto the plates. Also spot 5ul of 7.15 phage stock on each plate as a control. | ||
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'''V) Results''' | '''V) Results''' | ||
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+ | 2) Spot Test to Confirm Phage Viability | ||
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+ | : From the spot tests, 1 and 3C had a plaque at -4 (The plaque on -4 for 16 was incorrect, as we accidently added -2 to that spot). All plates have plaques for the 7.15 control. | ||
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[[File:BYUSuM5SpotTest.JPG|400px|center|link=]] | [[File:BYUSuM5SpotTest.JPG|400px|center|link=]] | ||
'''VI) Conclusion''' | '''VI) Conclusion''' | ||
- | + | It appears as though incubating the phage with mutagen for 1 hour is better than 3 hours and 16 hours; however, we cannot hold this to be terribly accurate because the control for 1 hour showed no plaques at -4, suggesting that there might be something wrong with the experiment. | |
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Revision as of 21:07, 23 July 2013
Small Phage July - August Notebook: Experiments
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7.17 Mutagen Concentration Test - Fifth Protocol
I) Purpose
II) Expected Outcome
III) Reagents Used
IV) Procedure 1) Applying the mutagen (7.17)
2) Spot Test to Confirm Phage Viability (7.19)
V) Results 2) Spot Test to Confirm Phage Viability
VI) Conclusion It appears as though incubating the phage with mutagen for 1 hour is better than 3 hours and 16 hours; however, we cannot hold this to be terribly accurate because the control for 1 hour showed no plaques at -4, suggesting that there might be something wrong with the experiment. |