As part of a research project i have obtained plant root samples that are currently frozen. I need to isolate bacterial DNA from the plant root, however i first need to remove the possible bacterial biofilm from the plant root as the bacterial DNA is of interest, not plant root DNA.
Any advice would be greatly appreciated
I would recommend you to thoroughly wash the root sample with double distilled water and collect the water. Inoculate the water any bacterial media according to your preference. From that culture you can isolate bacterial DNA without any fear of plant DNA contamination. But with this protocol you will have DNA of only loosely adhered bacterial population. To obtain DNA of strongly adhered bacterial population, you must cut the root sample into small pieces and homogenized in 85% NaCl solution. Then get rid of the root debris and use the supernatant to inoculate any bacterial media. From that media you can isolate DNA where will be no plant DNA contamination.
It works for me. But in my case I had fresh root sample. I think it will work for you too.
Thank you
Seana ni Fhearghail
What is your aim, what would you do with the DNA?
The suggestion above is fine but it has a very narrow relevance. Environmental microbiology does not work well with culturing techniques.
I agree with Debapriya Sarkar in this question but you can also use some commercially available kits, e.g., FastPrep DNA isolation kit for bacteria
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