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CharonY

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Everything posted by CharonY

  1. Spores don't propagate and can't be thermophilic per definitionem. They are thermo tolerant, however Regarding thermophilic pathogens, thermophilic Campylobacter species comes to my mind there are also some clostridia that have been classified as thermophilic.
  2. Actually I think I read somewhere that annealing of the DNA-templates (that is, forming of hydrogen bonds) is a (the?) rate limiting step in this reaction. At high temperatures these bonds are naturally less stable or easier disrupted. Of course, a host of protocols exist and depending on your templates it might not be essential at all. With sufficient vector and insert e.g. an 1-2 h incubation on the bench usually works. On the other hand I often do o/n incubations in thermos flasks in the fridge for blunt end ligation. Using the thermos flask the temperature decreases more slowly and supposedly increases efficiency...
  3. How about checking with a microbiologist at the nearest university? Setup is pretty straightforward. A standard test is to plate the bactaeria on an agar plate and put discs with antibiotics on it. Measure the zone of inhibition. Each discs contains a different AB (or concentration). Another standard test are MIC-tests (Minimum inhibition concentration). THese are more work. You need liquid medium, each containing rising concentrations of AB. You measure the optical density (OD) to determine which cocentration of a respective AB inhibits growth. Note that in many countries for safety reasons you are required to kill the bacteria after use, which requires autoclaving.
  4. Well there is actually quite a a bit research regarding introns and their function and it is now more or less known that they do carry information that will not directly transform to proteins (that is, get translated). And it ain't some central "dogma". The only dogma i am aware of is the information flow from DNA to mRNA to protein (although in case of reverse transcrpitase activity the information of mRNA might actually lead back to DNA). Same goes to so-called "junk" DNA. For quite a few of them ideas begin to develop what they might be fore (and as such the term junk is clearly a misnomer). Especially the research on small RNAs has led to the discovery of new regualtory roles (or in some cases, splicing functions). Check this for instance http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=14550267
  5. One can either add the substances to the (slightly cooled) agar while before pouring it into the petri dish). Alternatively there are antibiotic discs to test them. Or you can put a sterile filter disk on a plate with bacteria and drop around 10µl on it. For E. colis you might want to ask nearby universities, however you won't (or at least shouldn't) get Streptococci or Salmonella. Both species are usually human pathogenes and may not be cultivated outside specially equipped laboratories! Edit: forgot to add: most chemicals can be acquired by e.g.Sigma, but you may need an authorization to buy there (depends a bit on local laws).
  6. I suppose you mean a change in protein abundancy? That's theoretically possible but for a true quantification you got to use techniques like radioactive labelling or DIGE, or the usage of heavy nitrogen. With 2D PAGES as compared to 1D you get a better resolution and therefore you can visulize more proteins. And in fact there are plenty of papers regarding the proteome of fungi. I don't think that there are 2d gels only woth ribosomal proteins around though, since one of the strength of this technique is the visualisation of a lot of proteins in one gel. If one could separate the ribosmal proteins from all others (at the moment i'd have no clue how short of His-tagging and purifying them) an 1D PAGE should suffice.
  7. None of these qualify as new species. They are just mutant strains.
  8. Ah I think you are confusing stop codon with terminator here. An operon can contain several ORFs, but not the other way round. A gene ends with its stop codon
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