Which methods/chemicals can be use to separate the biofilm from a plastic bead. Tried many steps to extract complete DNA from biofilm attached to plastic beads used in a nitrifying bioreactor. Getting lots of DNA but not from the biofilm. The consortia in the bifilm were already known. The another issue is I have to remove the bacteria partially attached on the surface of biofilm, which are not contribute in biofilm formation. Please help
Hi,
To remove loosely adherent cells you should wash the plastic beads with sterilized buffered, may be a PBS (I think that two washing steps is enough). Then harvest the beads with biofilm by centrifugation at low rpm. You do not say anything about the diameter of this beads ? so centrifugation step may not be required.
To remove microorganisms from the surface you can cleaning beads with sonic energy (you must to optimize power and time). You can add to the cleaning solution some surfactant to improve the process efficiency. To ensure proper separation biofilm from plastic beads you can use light or fluorescence microscope.
good luck
Sonication in the presence of detergents may be useful in separating biofilms from plastic beads. You may need to optimize the detergent concentration and sonication frequency, and efficiency can be monitored by staining with fluorescent dye. “Getting lots of DNA but not from the biofilm” is a different problem; you need to try different DNA extraction methods. Usually, it is very difficult to extract DNA from biofilms. You can try soil DNA extraction kits.
Good luck
Hi, I am looking for the methods to extract biofilm from plastic beads taken from dentrifying bioreactor. The purpose is to Extract DNA for the qPCR. Can anyone please provide the protocol of the purpose?
Thanks
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