摘要
The polymerase chain reaction （PCR） has been a reliable and fruitful method for many applications in ecology. Nevertheless, unavoidable technical and instrumental require- ments of PCR have limited its widespread application in field situations. The recent development of isothermal DNA amplifica- tion methods provides an alternative to PCR, which circumvents key limitations of PCR for direct amplification in the field. Being able to analyze DNA in the pollen cloud of an ecosystem would provide very useful ecological information, yet would require a field-enabled, high-throughput method for this potential to be realized. Here, we demonstrate the applicability of the loop- mediated DNA amplification method （LAMP）, an isothermal DNA amplification technique, to be used in pollen analysis. Wedemonstrate that LAMP can provide a reliable method to identify species from the pollen cloud, and that it can amplify successfully with sensitivity down to single pollen grains, thus opening the possibility of field-based, high-throughput analysis.
The polymerase chain reaction （PCR） has been a reliable and fruitful method for many applications in ecology. Nevertheless, unavoidable technical and instrumental require- ments of PCR have limited its widespread application in field situations. The recent development of isothermal DNA amplifica- tion methods provides an alternative to PCR, which circumvents key limitations of PCR for direct amplification in the field. Being able to analyze DNA in the pollen cloud of an ecosystem would provide very useful ecological information, yet would require a field-enabled, high-throughput method for this potential to be realized. Here, we demonstrate the applicability of the loop- mediated DNA amplification method （LAMP）, an isothermal DNA amplification technique, to be used in pollen analysis. Wedemonstrate that LAMP can provide a reliable method to identify species from the pollen cloud, and that it can amplify successfully with sensitivity down to single pollen grains, thus opening the possibility of field-based, high-throughput analysis.
基金
Financial support was provided by Mexus, a collaborative program between the University of California and the Mexican National Council for Science and Technology, CONACyT
Further support provided by USDA- ARS funding
