Team:BYU Provo/Notebook/SmallPhage/Springexp/Period3/Exp/7.8 Mutagen Concentration Test - Forth Protocol
From 2013.igem.org
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: Plates were incubated upside down for 24 hours. | : Plates were incubated upside down for 24 hours. | ||
- | 3) | + | 3) Spot Test to Confirm Phage Viability (7.12) |
+ | |||
+ | : - Add 0.75mL of BL21 overnight into 8 test tubes | ||
+ | |||
+ | : - Add 7mL of x6 top agar to each tube and plate it on top of LB plates | ||
+ | |||
+ | : - Divide the plates into 6 quadrants with a sharpie. 4 plates are for the new dilution (next step) and 4 plates are for the old dilution series (step 2), each labeled with their respective mutagen concentration series. | ||
+ | |||
+ | : - Using the mutagenized phage stock from step 1 above, create a 1:10 dilution series of each mutagen concentration (0ul, 100ul, 200ul, and 500ul) from 0 to -4 using 90ul LB and 10ul phage in eppendorf tubes. | ||
+ | |||
+ | : - Spot 5ul of each eppendorf tube (from both new and old dilution) onto the plates. Also spot 5ul of 5.3 phage stock on each plate as a control. | ||
'''V) Results''' | '''V) Results''' | ||
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2) Titer to Determine Phage Concentration 1 | 2) Titer to Determine Phage Concentration 1 | ||
- | : Only the 0 plates from the 100ul, 200ul, and 500ul series showed plaques. All other plates had no plaques. The 0 200ul plate had the most plaques. | + | : - Only the 0 plates from the 100ul, 200ul, and 500ul series showed plaques. All other plates had no plaques. The 0 200ul plate had the most plaques. |
+ | |||
+ | 3) Spot Test to Confirm Phage Viability | ||
+ | |||
+ | : - The old dilution series plates showed plaques at 0 and 5.3. | ||
+ | |||
+ | : - The new dilution series plates showed the following plaques: | ||
+ | |||
+ | :: - 0ug: 5.3, 0 | ||
+ | |||
+ | :: - 100ug: 5.3, 0, -1 | ||
+ | |||
+ | :: - 200ug: 5.3, 0, -1, -2 | ||
- | 3 | + | :: -500 ug: 5.3, 0 |
'''VI) Conclusion''' | '''VI) Conclusion''' |
Revision as of 20:25, 15 July 2013
Small Phage July - August Notebook: Experiments
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7.8 Mutagen Concentration Test - Fourth Protocol
I) Purpose
II) Expected Outcome
III) Reagents Used
IV) Procedure 1) Applying the mutagen (7.8)
2) Titer to Determine Phage Concentration 1 (7.10)
3) Spot Test to Confirm Phage Viability (7.12)
V) Results 2) Titer to Determine Phage Concentration 1
3) Spot Test to Confirm Phage Viability
VI) Conclusion
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