20.109(F07): Phage by design, pt2: Difference between revisions
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==Protocols== | ==Protocols== | ||
===Part 1: M13.1 titers=== | ===Part 1: M13.1 titers=== | ||
The strains you transformed last time with M13K07 and M13.1 were inoculated into LB+Kan and grown overnight. If the two forms of the phage are equally robust, then the phage titer in the liquid surrounding the cells should be approximately the same. Ditto if the wild type strain, MG1655, and the MDS strain are equally capable of phage production. You will compare the PFU/ul of each culture to determine this. The protocol is identical to the one from [[20.109(F07): Agarose gel electrophoresis#Protocols| module 1]]. Since you don't know if the phage production will be improved or diminished by any of the variations, you will titer a wider-than-usual range of concentrations. | The strains you transformed last time with M13K07 and M13.1 were inoculated into LB+Kan and grown overnight. If the two forms of the phage are equally robust, then the phage titer in the liquid surrounding the cells should be approximately the same. Ditto if the wild type strain, MG1655, and the MDS strain are equally capable of phage production. You will compare the PFU/ul of each culture to determine if this is true. The titering protocol is identical to the one from [[20.109(F07): Agarose gel electrophoresis#Protocols| module 1]]. Since you don't know if the phage production will be improved or diminished by any of the variations, you will titer a wider-than-usual range of concentrations. | ||
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You should also include a no phage control plate for confidence that the number of plaques you see next time arise as a result of your experiment. | You should also include a no phage control plate for confidence that the number of plaques you see next time arise as a result of your experiment. | ||
===Part 2: Phage deposition onto patterned slide=== | ===Part 2: Phage deposition onto patterned slide=== | ||
Revision as of 12:13, 2 November 2007
Introduction
Protocols
Part 1: M13.1 titers
The strains you transformed last time with M13K07 and M13.1 were inoculated into LB+Kan and grown overnight. If the two forms of the phage are equally robust, then the phage titer in the liquid surrounding the cells should be approximately the same. Ditto if the wild type strain, MG1655, and the MDS strain are equally capable of phage production. You will compare the PFU/ul of each culture to determine if this is true. The titering protocol is identical to the one from module 1. Since you don't know if the phage production will be improved or diminished by any of the variations, you will titer a wider-than-usual range of concentrations.
| phage = M13K07, strain = MG1655 | phage = M13K07, strain = MDS | phage = M13.1, strain = MG1655 | phage = M13.1, strain = MDS | |
| 10-6 | ||||
| 10-8 | ||||
| 10-10 |
You should also include a no phage control plate for confidence that the number of plaques you see next time arise as a result of your experiment.
Part 2: Phage deposition onto patterned slide
DONE!
For next time
- water
- how to more densely pack nanowires
