Enhancing PCR for Prey DNA in Antarctic Krill Stomachs
Author Information
Author(s): Hege Vestheim, Simon N. Jarman
Primary Institution: Department of Biology, University of Oslo; Australian Antarctic Division
Hypothesis
Can blocking primers improve the PCR amplification of rare prey DNA in the presence of dominant predator DNA?
Conclusion
The study presents a method that effectively amplifies prey DNA while blocking predator DNA in PCR mixtures.
Supporting Evidence
- The method reduced predator amplicons to undetectable levels even when predator template was present in 1000-fold excess.
- Adding blocking primers allowed for the successful amplification of prey DNA.
- 111 clones from 13 different krill stomachs revealed 36 different sequences.
Takeaway
Scientists found a way to make sure they can see the food that krill eat by blocking the DNA from the krill itself when they look for the DNA of the food.
Methodology
The study involved designing blocking primers to prevent the amplification of predator DNA while allowing prey DNA to be amplified in PCR.
Potential Biases
The blocking primers may inadvertently block other non-target species' DNA.
Limitations
The method may not completely eliminate the risk of amplifying other species' DNA, and the complete sequence coverage of potential prey was not available.
Participant Demographics
The study focused on Antarctic krill and their dietary analysis.
Digital Object Identifier (DOI)
Want to read the original?
Access the complete publication on the publisher's website