I have a two-pronged plan to get a phage strain that gives good enough plaques for my GTA-as-vaccine experiments.
I obtained reasonable titers of two phages, 'Titan' and 'Saxon'. I'll invest a couple of weeks to see if I can get better and more reproducible plaques with either of these. The genome sequences of these phages are not closely related.
First, improve the plaquing conditions: The researcher who isolated the phages recommends using for the lawn cells that have been grown photosynthetically to a high density, He also suggested trying a lower top-agar concentration. I'll play around with these and other variables to see if I can get better plaques.
Second, use artificial selection to get a better strain of phage: I'll pick the few best-looking plaques of each of my two phages and plate the phage they contain in new lawns. From those new lawns I'll again pick the best-looking plaques, and plate their phage in new lawns. Etc. Maybe I'll introduce a bit of UV mutagenesis along the way.
The first step will be to make fresh lysates of these phages. The lawns I made before are too old, so I'll grow up some cells for lawns today and tomorrow I'll retiter the lysates. On Friday I can pick plaques from these lawns and make plate lysates. If there's a plate with near-confluent plaques I can use it directly to make a plate lysate. (10^7 or 10^8 pfu/ml, and I have maybe 5 µl so at best I can get plates with 5 x 10^4 or 5 x 10^5 plaques. The latter might be enough to get a good lysate. There are small volumes (50-100 µl?) of the original lysates in the lab upstairs, so maybe I'll sue these. Or Maybe I should save these until I've improved the plaquing conditions.
- Home
- Angry by Choice
- Catalogue of Organisms
- Chinleana
- Doc Madhattan
- Games with Words
- Genomics, Medicine, and Pseudoscience
- History of Geology
- Moss Plants and More
- Pleiotropy
- Plektix
- RRResearch
- Skeptic Wonder
- The Culture of Chemistry
- The Curious Wavefunction
- The Phytophactor
- The View from a Microbiologist
- Variety of Life
Field of Science
-
-
-
Political pollsters are pretending they know what's happening. They don't.5 weeks ago in Genomics, Medicine, and Pseudoscience
-
-
Course Corrections6 months ago in Angry by Choice
-
-
The Site is Dead, Long Live the Site2 years ago in Catalogue of Organisms
-
The Site is Dead, Long Live the Site2 years ago in Variety of Life
-
Does mathematics carry human biases?4 years ago in PLEKTIX
-
-
-
-
A New Placodont from the Late Triassic of China5 years ago in Chinleana
-
Posted: July 22, 2018 at 03:03PM6 years ago in Field Notes
-
Bryophyte Herbarium Survey7 years ago in Moss Plants and More
-
Harnessing innate immunity to cure HIV8 years ago in Rule of 6ix
-
WE MOVED!8 years ago in Games with Words
-
-
-
-
post doc job opportunity on ribosome biochemistry!9 years ago in Protein Evolution and Other Musings
-
Growing the kidney: re-blogged from Science Bitez9 years ago in The View from a Microbiologist
-
Blogging Microbes- Communicating Microbiology to Netizens10 years ago in Memoirs of a Defective Brain
-
-
-
The Lure of the Obscure? Guest Post by Frank Stahl12 years ago in Sex, Genes & Evolution
-
-
Lab Rat Moving House13 years ago in Life of a Lab Rat
-
Goodbye FoS, thanks for all the laughs13 years ago in Disease Prone
-
-
Slideshow of NASA's Stardust-NExT Mission Comet Tempel 1 Flyby13 years ago in The Large Picture Blog
-
in The Biology Files
Not your typical science blog, but an 'open science' research blog. Watch me fumbling my way towards understanding how and why bacteria take up DNA, and getting distracted by other cool questions.
No comments:
Post a Comment
Markup Key:
- <b>bold</b> = bold
- <i>italic</i> = italic
- <a href="http://www.fieldofscience.com/">FoS</a> = FoS