Impact of Primer Selection on Microbial Community Profiles
Author Information
Author(s): Kumar Purnima S., Brooker Michael R., Dowd Scot E., Camerlengo Terry
Primary Institution: The Ohio State University
Hypothesis
The hypervariable region targeted for sequencing and primer degeneracy play important roles in influencing the composition of 16S pyrotag communities.
Conclusion
The hypervariable region targeted by the primer significantly influences the profile of complex microbial communities generated by pyrosequencing.
Supporting Evidence
- Greater numbers of species were detected by pyrosequencing than by Sanger sequencing.
- Rare taxa constituted nearly 6% of each pyrotag community and less than 1% of the Sanger sequencing community.
- The communities generated by degenerate and non-degenerate primers did not demonstrate significant differences.
Takeaway
This study looked at how different primers used in DNA sequencing can change the picture we get of tiny living things in our mouths. It found that using the right primers helps us see a more complete picture of these communities.
Methodology
Subgingival plaque samples were collected from smokers with chronic periodontitis and analyzed using Sanger sequencing and pyrosequencing with four different primer pairs.
Potential Biases
Potential bias from primer selection and sequencing methods could affect the representation of certain taxa.
Limitations
The study was limited to a small sample size of smokers with chronic periodontitis, which may not represent other populations.
Participant Demographics
10 current smokers with generalized moderate to severe chronic periodontitis.
Statistical Information
P-Value
p<0.001
Statistical Significance
p<0.01
Digital Object Identifier (DOI)
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