Team:BYU Provo/Notebook/SmallPhage/Summerexp/8.16 Modeling phage plaque size
From 2013.igem.org
(Difference between revisions)
Line 65: | Line 65: | ||
* Propagated T1 by infection 1mL E coli B and 4mL of LB with 10uL of stock phage. Complete clearage was observed in two days before purification via centrifuge (3000rpm for 5min) and chloroform (100uL of chloroform to 1mL of lysis). This stock of T1 is labeled T1P. | * Propagated T1 by infection 1mL E coli B and 4mL of LB with 10uL of stock phage. Complete clearage was observed in two days before purification via centrifuge (3000rpm for 5min) and chloroform (100uL of chloroform to 1mL of lysis). This stock of T1 is labeled T1P. | ||
- | * Dilution series -2 through -12 was performed for | + | * Dilution series -2 through -12 was performed for T1P. 5uL of each dilution sample was spotted onto plates overlaid with 0.5mL of E coli B and 5mL of x1 agar. |
3) Spot test for T2 from stock (8.20) | 3) Spot test for T2 from stock (8.20) | ||
Line 73: | Line 73: | ||
4) Preliminary Titer (9.8) | 4) Preliminary Titer (9.8) | ||
- | * Based on results from spot test for | + | * Based on results from spot test for T1P and T2, titer was performed for T1P 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. |
+ | |||
+ | 5) Repeated plating for T1 and T2 (9.9) | ||
+ | |||
+ | * Preliminary titer revealed that T1P at -7 and T2 at -4 have adequate concentrations for plating. Plating procedures were the same as those recorded on 8.16. The only difference is the initial OD 600 reading of E coli B liquid culture overnight and subsequent dilutions: the average between two OD 600 readings was 0.513, but dilution was performed right after to adjust it to 0.5. | ||
'''V) Results''' | '''V) Results''' |
Latest revision as of 06:56, 10 September 2013
| |||||||||||||||||||||||||||||||||||||||||||
|
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)
5) Repeated plating for T1 and T2 (9.9)
V) Results 1) Plating
2) T1 and T2 spot test
|