Team:BYU Provo/Notebook/SmallPhage/Summerexp/8.16 Modeling phage plaque size
From 2013.igem.org
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* To see a statistical significance in plaque sizes between different phages, many plaques are needed. Thus 10 plates were used for T1, T2, T3, and T7. Because T4 form much smaller plaques, only 3 plates were used. | * To see a statistical significance in plaque sizes between different phages, many plaques are needed. Thus 10 plates were used for T1, T2, T3, and T7. Because T4 form much smaller plaques, only 3 plates were used. | ||
- | * Experimental procedures are as follow: 0.5mL of E coli BL21 (OD 0.5, adjusted after initial measurement of 0.558) was infected with 20uL of respective phage dilution samples for 10 minutes. 5mL x1 top agar was then added to each test tube, and the content was plated. The plates were incubated for 24 hours. | + | * Experimental procedures are as follow: 0.5mL of E coli BL21 (OD 0.5, adjusted after initial measurement of 0.558) was infected with 20uL of respective phage dilution samples for 10 minutes. 5mL x1 top agar was then added to each test tube, and the content was plated. The plates were incubated at 37 Celsius for 24 hours. |
2) Propagation and spot test of T1 (8.12-8.20) | 2) Propagation and spot test of T1 (8.12-8.20) | ||
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* Dilution series -2 through -7 was performed for T2 from stock. 5uL of each dilution sample was spotted onto plates overlaid with 0.5mL of E coli B and 5mL of x1 agar. | * Dilution series -2 through -7 was performed for T2 from stock. 5uL of each dilution sample was spotted onto plates overlaid with 0.5mL of E coli B and 5mL of x1 agar. | ||
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+ | 4) Preliminary Titer (9.8) | ||
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+ | * Titer was performed for T1 at -6, -7, and -8; and T2 at -2, -3, and -4. Specifically, 0.5mL of E coli B liquid culture overnight was infected with 20uL of respective phage sample. After 15 minutes of incubation, 5mL of x1 top agar (not precise) was added to each test tube, and the content was plated. The plates were incubated at 37 Celsius for 20 hours. | ||
'''V) Results''' | '''V) Results''' |
Revision as of 00:16, 9 September 2013
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8.16 Modeling phage plaque size - Experiment One
I) Purpose
II) Expected Outcome
III) Reagents Used
IV) Procedure 1) Plating (8.16)
2) Propagation and spot test of T1 (8.12-8.20)
3) Spot test for T2 from stock (8.20)
4) Preliminary Titer (9.8)
V) Results 1) Plating
2) T1 and T2 spot test
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