Team:BYU Provo/Notebook/SmallPhage/Summerexp/8.2 Modeling phage plaque size
From 2013.igem.org
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- | <font size="5"> ''' | + | <font size="5"> '''8.2 Modeling Phage Plaque Sizes - Preliminary Experiments''' </font> |
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* To start modeling phage plaque sizes against several variables including, but not limited to phage particle size, agar concentration, and bacterial concentration. | * To start modeling phage plaque sizes against several variables including, but not limited to phage particle size, agar concentration, and bacterial concentration. | ||
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+ | * To determine variables within modeling and make preparation for actual experiments | ||
'''II) Expected Outcome''' | '''II) Expected Outcome''' | ||
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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. | * Dilution series from -2 to -7 was performed for T1, T2, T3, T4, and T7. 5ul of each dilution was spotted onto respective plates. | ||
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+ | 4) Preliminary titer test (8.10) | ||
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+ | * Titer was performed for (...) dilution samples. | ||
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+ | * Specifically, 0.5mL of E coli BL21 was infected with 20uL of respective phage dilution samples for approximately 15 minutes. x1 top agar was then added to each test tube, and the content was plated. | ||
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+ | * Plates were incubated for approximately 24 hours. | ||
'''V) Results''' | '''V) Results''' | ||
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'''VI) Conclusion''' | '''VI) Conclusion''' | ||
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+ | * There are multiple variable intrinsic to the design of the modeling experiment that we'll have to control for. These include phage speciation (particle size, infection rate, lysis rate, etc), agar concentration, adsorption time, bacterial density, and incubation time. | ||
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+ | * For actual experiments that model phage plaque size against phage particle size we will have to control for all variable apart from phage speciation. This implies that we will have to make top agar solution with a volumetric flask, measure adsorption time, use OD to determine bacterial density, and measure incubation time. It will be easiest if we can standardize adsorption time, bacterial density, and incubation time so that we can compare results between different experiments. | ||
Revision as of 22:47, 10 August 2013
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8.2 Modeling Phage Plaque Sizes - Preliminary Experiments
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)
4) Preliminary titer test (8.10)
V) Results 1) E coli strain preparation
2) Phage viability/infection test
3) Phage titer determination
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