Extracting nucleic acids from activated sludge which reflect community population diversity |
| |
Authors: | Simon J McIlroy Kate Porter Robert J Seviour Daniel Tillett |
| |
Institution: | (1) Biotechnology Research Centre, La Trobe University, P.O. Box 199, Bendigo, VIC, 3552, Australia;(2) Biota Holdings Limited, 10/585 Blackburn Road, Notting Hill, VIC, 3168, Australia |
| |
Abstract: | Critical to most studies in molecular microbial ecology is the application of DNA/RNA extraction methods which can reveal
the true level of population biodiversity present in samples from the community under investigation. Activated sludge communities
have been studied extensively using molecular methods, but rarely have the nucleic acid isolation methods applied been assessed
for their ability to achieve this. This study compares eight published RNA and DNA extraction protocols and one commercially
available DNA isolation kit for their capacity to provide high quality nucleic acids that reflect the community composition.
Each method was assessed on the basis of nucleic acid yield, purity and integrity, and the ability to provide PCR amplifiable
RNA and DNA from known marker populations that varied in their resistance to nucleic acid extraction. Only three consistently
provided DNA from each of the marker populations known to be present in the samples from fluorescence in situ hybridisation
analysis. The failure of the other methods emphasises the need to validate all DNA/RNA extraction protocols. It is recommended
that several validated extraction methods be used and the extracts pooled to further minimise any risk of bias. |
| |
Keywords: | |
本文献已被 SpringerLink 等数据库收录! |
|