Abstract
The comparison of the bacterial profile of intracellular (iDNA) and extracellular DNA (eDNA) isolated from cow rumen content stored under different conditions was conducted. The influence of rumen fluid treatment (cheesecloth squeezed, centrifuged, filtered), storage temperature (RT, -80 °C) and cryoprotectants (PBS-glycerol, ethanol) on quality and quantity parameters of extracted DNA was evaluated by bacterial DGGE analysis, real-time PCR quantification and metabarcoding approach using high-throughput sequencing. Samples clustered according to the type of extracted DNA due to considerable differences between iDNA and eDNA bacterial profiles, while storage temperature and cryoprotectants additives had little effect on sample clustering. The numbers of Firmicutes and Bacteroidetes were lower (P
Original language | English |
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Pages (from-to) | 80-84 |
Number of pages | 5 |
Journal | Anaerobe |
Volume | 29 |
DOIs | |
Publication status | Published - 12 Oct 2013 |
Event | 8th International Symposium on Anaerobic Microbiology (ISAM 8): No oxygen - still vigorous - Innsbruck, Austria Duration: 12 Jun 2013 → 15 Jun 2013 |
Keywords
- Intracellular DNA
- Extracellular DNA
- Storage conditions
- Rumen fluid
- Bacterial diversity
- Firmicutes
- Bacteroidetes
- PCR-DGGE
- Q-PCR
- Metabarcoding