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Detaching Biofilms


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7 replies to this topic

#1 autoclave

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Posted 01 May 2009 - 04:02 AM

Hi Everyone !

I need to detach a biofilm from a surface which will then be processed to extract DNA. I am just wondering if sonication will be enought to remove all attached bacteria. The surface the biofilm will be attached to is a thin clay tablet. Any ideas?

Cheers

#2 GeorgeWolff

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Posted 01 May 2009 - 02:51 PM

Why not remove it with a tool - like a sterile spatula?

#3 autoclave

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Posted 02 May 2009 - 05:04 AM

Why not remove it with a tool - like a sterile spatula?


I believe that removing it with a sterile spatula would still perhaps leave bacteria attached to the surface. In very thin biofilms, on the other hand, I guess it would not work (?).

#4 phage434

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Posted 02 May 2009 - 05:30 AM

I would include a sample of the clay tablet in the lysis reaction. Chemistry will remove things that mechanical disruption will not. A phenol/chloroform extraction would likely remove most anything organic.

#5 autoclave

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Posted 02 May 2009 - 06:47 AM

I would include a sample of the clay tablet in the lysis reaction. Chemistry will remove things that mechanical disruption will not. A phenol/chloroform extraction would likely remove most anything organic.

thanks for your reply. should the phenol/chloroform extraction of the biofilm interfere with subsequent DNA extraction from it using e.g., a DNA isolation kit?

cheers

#6 hanming86

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Posted 06 May 2009 - 06:56 PM

i would go with georgewolff's method of using a spatula to scrap the biofilm out. It should get like most of the thing. even if it's a thin biofilm u will still get something. It's kinda like u know scraping bacteria off agar plate. the more distance u scrap the "thicker" the cell mass get. Same theory .
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#7 archercr

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Posted 12 March 2010 - 08:26 AM

I've read of scraping with a spatula, but in my protocol, we sonicate in a water bath. You place a steel tray inside the water bath and the plate inside the dry tray. the vibrations from the water transfer through the tray to the plate, causing the bacteria to loosen.

#8 microgirl

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Posted 12 March 2010 - 12:06 PM

I just got an email from MoBio about their new kit:
http://www.mobio.com...es/biofilm.html
I don't do biofilms, so I didn't look into it, but maybe it has some suggestions.




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