I am extracting DNA from soil sample based on Zhou's conventional method but I am facing problems with the humic acids. The humic acids will decrease the purity of DNA and thus, inhibits the downstream process. Is there any method to remove this?
Dear you just add active charcol during your lysis steps and also you should treat PEG-8000 precipitation after that SDS lysis step. All the best
We tried a couple of modifications to Zhou's protocol, to make it work for compost DNA. As Yugal stated, we found using PEG to preciptate helped (LaMontagne et al. 2002). After PEG precipitation go immediately into CTAB/choloroform extraction and you should see the humic acids go into the CTAB interface.
If you are still looking for protocol suggestions we have used the method from Sagova-Mareckova (2008). Seems to deliver reliably amplifiable DNA.
I tried runing and electrophoresis with 2% PVP on borate buffer and cut the band. Works fine, also I could amplified 16Srna gene from the sample, but I lost to mucch DNA on purification proces (80%)
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