Plasmid Extraction from P. Aeruginosa smears in Gel! (View forum version)



tihong10

Posted 11 April 2013 - 01:13 AM

Hi, I electroporated P. aeruginosa 14 with a plasmid and when I tried to view the plasmid extraction in a gel it came out as a smear. Is there some big issue I am not aware of when plasmid extracting from PA? Is my plasmid being degraded somehow?

I wasn't sure how much detail to put but if more is needed I will provide it.

Thanks!

pito

Posted 12 April 2013 - 04:00 AM

could be so many things...
perhaps putting a picture of the gel here could help?

it could be that you somehow broke the plasmid or that you are seeing non plasmid dna...

tihong10

Posted 16 July 2013 - 11:48 PM

Hello,

I wanted to bump this post back up...

Here is a gel image of the P. aeruginosa 14 (transformant containing a recombinant plasmid) plasmid extraction that I performed using a Plasmid miniprep kit (using the optional denaturation buffer).

Whenever I perform a P.E. on PA14 I always get this kind of smear. Can anyone shed some light on this smearing pattern that keeps appearing and possibly a way I can cleanly extract a plasmid from the PA14 bacteria?

Thank you,
Ted