OK, I've done the third of the experiments I planned here, and the results cleanly show that nothing interesting is going on (at least nothing interesting that warrants investigation).
I wanted to know if the peculiarities I had long-ago noticed in DNA of competent cells were reproducible. So yesterday I grew some wildtype cells in rich medium. Some of them I collected during exponential growth ('log phase', cell density about 2 x 10^9 cells/ml), some when the culture was approaching its final density ('stationary phase', about 10^10 cells/ml), and some I transferred to the competence-inducing starvation medium while they were in log phase. I prepared chromosomal DNA from all three treatments, and then incubated it either in the standard DNA buffer TE (10 mM Tris pH 8, 1 mM EDTA) or in the mung bean nuclease buffer that had previously given anomalous results for competent-cell DNA. DNA in the mung bean nuclease buffer sat at room temperature for about an hour, with and without being heated to 65 °C for 10 minutes. Then I ran all the DNAs in a gel to check their condition.
You can see that there's no significant differences between the effects of the treatments on the different DNAs. The competent cell DNA is in slightly shorter fragments (the biggest marker band is 29 kb), but the difference between it and the other DNAs isn't changed by the MBN buffer. Heating the DNAs in the buffer causes the usual streaking of the bands (caused by having a lot of high-molecular-weight DNA running at the same position) to be a bit blurry, but again that applies equally to all three DNAs.
So I can put this old result out of my mind, and get to work on my planned competence and phage recombination assays.
- 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