Authors:
Sharon Yu Ling Lau, Chai Fung Pui, Yasuyuki Hashidoko and Lulie Melling
Book:
Proceedings of the 14th International Peat Congress
Venue:
Stockholm
Keywords:
dna-extraction, humic-substances, tropical-peat-soil
Documentfile:
Lau et al 2012: Comparison of Methods for Total Community DNA Extraction from Tropical Peatland
Summary:
Theme IX. Tropical peatlands
SUMMARY
Soil DNA extraction is usually complicated by the co-purified contaminants especially humic substances, which subsequently inhibit post-extraction analyses such as PCR-DGGE. Conventional methods employing sodium dodecyl sulphate (SDS) and lysozyme-based treatment yield better DNA quantity but higher amount of humic substances are also being co-extracted together with the total DNA. Nucleospin (Macherey-Nagel, Germany) obtained PCR-amplifiable DNA with purity of A260nm/A230nm ratio of 1.70 and humic acid level measuring at 0.0041 (A320nm). PCR amplification was also improved by using skimmed milk and Pfu DNA polymerase.