Team:BYU Provo/Notebook/SmallPhage/Summerexp/8.2 Modeling phage plaque size
From 2013.igem.org
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2) Phage viability/infection test (8.5) | 2) Phage viability/infection test (8.5) | ||
- | + | * Prepare 2 plates for a spot test using E. coli BL21 by adding 0.5mL overnight into a tube, mixing it with 5mL of x1 top agar, and then plating it. Repeat for E. coli B and E. coli W3110. | |
- | + | * Spot 5ul each of T1, T3 30, T3 14, T5, and T7 (7.19 10uL stock) onto one plate of each bacteria. Spot T2 10, T2 30, T4 (5E9 pfu/mL as determined by the large phage group), Phi X 174 20, and Phi X174 40 onto the other plate of each bacteria. | |
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+ | 3) Phage titer determination (8.7) | ||
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+ | * Five plates, labeled T1, T2, T3, T4, and T7, was prepared for spot tests using E. coli B by adding 0.5mL overnight into a tube, mixing it with 5mL of x1 top agar, and then plating it. | ||
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+ | * Dilution series from -2 to -7 was performed for T1, T2, T3, T4, and T7. 5ul of each dilution was spotted onto respective plates. | ||
'''V) Results''' | '''V) Results''' | ||
- | 1) E coli K12 did not survive on plate or in liquid culture overnight. So we decided to viability/infection of phage with E coli B, BL21, and W3110 instead. | + | 1) E coli strain preparation |
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+ | * E coli K12 did not survive on plate or in liquid culture overnight. So we decided to viability/infection of phage with E coli B, BL21, and W3110 instead. | ||
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+ | 2) Phage viability/infection test | ||
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+ | * E coli BL21 supports the growth of bacteriophage T2, T3 30, and T7. On the other hand, E coli B and E coli W3110 support bacteriophage T1, T2 30, T3 30, T4, and T7. Other phage stocks either do not infect the bacteria strains tested or have died during storage in the fridge. | ||
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+ | * Because E coli B is the strain routinely used for the Large Phage Group's experiments, we decided to base our modeling on E coli B. | ||
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'''VI) Conclusion''' | '''VI) Conclusion''' |
Revision as of 22:38, 7 August 2013
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7.29 Mutagen Concentration Test - Sixth Protocol
I) Purpose
II) Expected Outcome
III) Reagents Used
IV) Procedure 1) E coli strain preparation (8.2-8.5)
2) Phage viability/infection test (8.5)
3) Phage titer determination (8.7)
V) Results 1) E coli strain preparation
2) Phage viability/infection test
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