We are filtering river water, of different salinities. With some samples, even when spiked with DNA of the species in question, we get no amplification. We've tried environmental mastermix.
You may try the phenol purification method.
Moore D, Dowhan D. Purification and concentration of DNA from aqueous solutions. Current protocols in pharmacology. 2001 May:A-3C.
http://onlinelibrary.wiley.com/doi/10.1002/0471140864.psa04es13/pdf
Just a suggestion.
http://www.aslo.org/lomethods/free/2004/0365.pdf
Optimization of DNA extraction for quantitative marine bacterioplankton community analysis
Hi there,
You can try using the meta-g-nome Epicenter kit (water, soil, etc..,). I have successfully isolated inhibitor free metagenomic DNA using meta-genome kit.
http://www.epibio.com/applications/nucleic-acid-purification-extraction-kits/dna-purification-genomic/meta-g-nome-dna-isolation-kit
Alternatively you can isolate DNA using routine protocol and electro elute the DNA from gel.
Dinesh
A few suggestions-
Dry your pellet in a speed vac and was it a few times with 70% EtOh, then resuspend in water.
Alternatively, a little Bovine Serum Albumin added to the mastermix can work well (it's also a bit imprecise).
- Badges
- Science method