Difference between revisions of "20.109(F07): Growth of phage materials"
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(New page: {{Template:20.109(F07)}} ==Introduction== ==Protocols== In advance of this lab, a bacterial host (namely XL1-blue) was infected with the modified M13 phage called "3-12." Though the precis...) |
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==Introduction== | ==Introduction== | ||
==Protocols== | ==Protocols== | ||
| − | In advance of this lab, a bacterial host (namely XL1-blue) was infected with the modified M13 phage called "3-12." Though the precise details of the 3-12 modification are still unpublished, | + | In advance of this lab, a bacterial host (namely [http://www.stratagene.com/products/displayProduct.aspx?pid=287| XL1-blue]) was infected with the modified M13 phage called "3-12." Though the precise details of the 3-12 modification are still unpublished, it was isolated from a screen for changes in the M13 p8 that enable the phage to bind a metal (Iridium). Today you will harvest the phage from the supernatant of the infected bacterial culture and then titer it. |
===Part 1: Phage purification=== | ===Part 1: Phage purification=== | ||
| + | #Spin 2 eppendorf tubes with infected cells in a room temperature microfuge, 1 minute at full speed. | ||
| + | #Remove the supernatant to clean fresh eppendorf tubes. | ||
| + | #Use your P1000 to measure the volume, then add a 1/6th volume of a 20% PEG-8000/2.5M NaCl solution. | ||
| + | #Invert to mix then incubate on ice 60 minutes. | ||
| + | #Spin in a room temperature microfuge, 15 minutes at full speed. | ||
Revision as of 14:54, 12 July 2007
Contents
Introduction
Protocols
In advance of this lab, a bacterial host (namely XL1-blue) was infected with the modified M13 phage called "3-12." Though the precise details of the 3-12 modification are still unpublished, it was isolated from a screen for changes in the M13 p8 that enable the phage to bind a metal (Iridium). Today you will harvest the phage from the supernatant of the infected bacterial culture and then titer it.
Part 1: Phage purification
- Spin 2 eppendorf tubes with infected cells in a room temperature microfuge, 1 minute at full speed.
- Remove the supernatant to clean fresh eppendorf tubes.
- Use your P1000 to measure the volume, then add a 1/6th volume of a 20% PEG-8000/2.5M NaCl solution.
- Invert to mix then incubate on ice 60 minutes.
- Spin in a room temperature microfuge, 15 minutes at full speed.
Part 2: Titering phage
DONE!
