Does my bacteria have a plasmid?
Posted 27 September 2004 - 12:54 AM
Posted 12 October 2004 - 02:40 AM
Posted 14 October 2004 - 05:09 AM
By the way, can you tell me which kit is better about it?
Posted 18 October 2004 - 01:23 AM
All protocols i know have been designed for coli which is gram -. i don't think it should make much difference expect you'd probably have to add a step to break down cell wall - lysosyme might do it. also, if your cells clump that might also be a problem.
i would try searching through Google or Web of Science for articles that have isolated plasmids from micrococcus, and look at their methods.
About the companies - I use the "Qiaprep spin miniprep kit", from Qiagen for isolating plasmids from vibrios (which are gram-negative) and it works great (though it was designed for E.coli). Roche is also very good but i have never tried it for anything other than coli. other companies of good reputation which probably have kits are Sigma and Promega. All these companies have technical assitance departments about their products and you can ask them about gram plus. usually its best to find their representatives in your own country and work through them.
Posted 24 October 2004 - 03:42 AM
Especially cause it's cheaper
Posted 16 November 2004 - 08:31 AM
One sugestion is using tranditional method and then try to further isolate plasmid from a crude DNA preparation.
Posted 25 May 2009 - 06:28 PM
It would not harm to try out the commercial one first.
But then again traditional method seems to be safer in this situation.
Posted 27 May 2009 - 02:37 AM
You can try to design primers to detect that part of the plasmid.
I've never tried it but it might work.