Sharon Yu Ling Lau, Chai Fung Pui, Yasuyuki Hashidoko and Lulie Melling
Proceedings of the 14th International Peat Congress
dna-extraction, humic-substances, tropical-peat-soil
Theme IX. Tropical peatlands
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.