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Best method for extraction on low concentration environmental DNA?

DNA Extraction Molecular Low-concentration

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#1 firefly2280

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Posted 05 November 2012 - 08:43 AM

Hi all

I could really do with some help. I am swabbing environmental sites, ie floors, surfaces (a 10cm3 area) and trying to extract DNA. Needless to say, Im not having a lot of luck, I think quantities are very low and inhibitors are present.

I have tried Qiagen colums with a bead-beating step first and also Phenol:chloroform. The column based method was better but neither give me a lot of DNA.

Does anyone have any ideas about increasing my yield please?

thanks

#2 El Crazy Xabi

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Posted 05 November 2012 - 07:54 PM

Lucky you have some... Posted Image I've been struggling with some samples that barely have any cells... and using 10 g of sample Posted Image

If inhibitors are a problem you may want to try some MoBIO kits, they use a patented inhibitor removal technology (as they call it) that works pretty well with difficult samples.

Don't know if it would help:
http://www.mobio.com...dna-from-swabs/

And I think they send kit samples to test

Edited by El Crazy Xabi, 05 November 2012 - 07:55 PM.


#3 firefly2280

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Posted 07 November 2012 - 03:03 AM

Thanks

I tried the Mobio kits, maybe I should give them another go! Thanks for your help and good luck with your work...!





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