I need plasmid extraction protocol from S. aureus without using a commercially available kit.
This may be the ticket: http://www.sciencedirect.com/science/article/pii/0003269781904735
The STET prep (boiling) cited was a long standard prep for many folks for routine minipreps in E.coli, I would note that the insoluble clot is generally just removed with a toothpick as oppose to the centrifugation mentioned, but have never tried it with G+ bugs. The key to a Staph preps is the lysozyme (or lysostaphin), in my experience (although not S. aures specifically) any variation of alkaline lysis using a Solution I that contains a liberal amount of lysozyme and proteinase K works. I toss in the proteinase K to combat nucleases prior to organic extraction, it may well be unnecessary. The cited lactobacilli protocol is such a protocol, it should work. They do two rounds of the isopropanol precipitation in that protocol (one prior to organic extraction), I do one.
Hi,
Try this, it works for most gram positive bacteria (pdf file attached).
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC239314/pdf/aem00166-0025.pdf
Regards
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