colony pcr question
L.A.M.Buisman at cable.A2000.nl
Fri Jan 7 15:01:41 EST 2000
Steven Projan wrote:
> Not all gram positives are created equal and lysozyme can often be an inappropriate choice to crack open the cell wall. For example Staphylococci are farily well imprevious to most muramidases but lysostaphin is quite effective, especially against the coagulase positive staph (e.g. S. aureus). Another useful enzyme is mutanolysin which has some activity against staphylococci and is an important tool in breaking open the enterococci (sometimes used in combination with lysozyme). Ogften doing a litle homework goes a long - check out volume 204 of Methods in Enzymology for some of this information as well publications describing work on the bacterium of interest.
I used a so-called "Mini Bead Beater". A small, fast shaker that holds 2ml. polypropylene screw-capped tubes with zirkonium beads. Cell-walls break up, DNA shearing is negligable.
I forgot which manufacturor.
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