Hi everyone...i just want to ask..i want to did plasmid extraction of various species of staphylococcus by using plasmid kit from Qiagen... i read many journal suggested about using lysostaphin instead of lysozyme..unfortunately, i deal with large sample (around 200 sample) and lysostaphin is very expensive. So, is it anyone experience did plasmid extraction from staphylococcus species with lysozyme and what suitable concentration of lysozyme for lysis?..Thank you..
Dear Abdul Rahim
You can using lysozyme in extraction of plasmid from staphylococcus
Best regrades
Thank you for your suggestion...is it you know the best concentration of lysozyme for staphylococcus plasmid extraction?
Dear Abdul Rahim,
You can use non commercial method that work very well to harvest a pure plasmid preparation. I used many methods on Staph and Streptococcus, and you don't need to use lysostaphin du to economical reason. Instead of you can incubate your bacterial culture in media suplemented with 0.5% Glycine to prevent cell wal synthetis. and your lysis will work by digesting with 50 mg/ml of lysozyme and 25% sucrose followed by alacaline extraction.
You can also read this publication :
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC168154/pdf/623527.pdf
Good luck
I suggest you use the method suggested above by Hafid Soualhine. It worked very well for me too It is alkaline extraction and not alacaline extraction as written
- Badges
- Science topic