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Plasmid Extraction from P. Aeruginosa smears in Gel!

plasmid extraction pa14 smear gel

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2 replies to this topic

#1 tihong10

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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!

#2 pito

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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...

If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#3 tihong10

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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





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