Simple colony PCR procedure for the filamentous actinobacteria Frankia
Molecular analysis of the filamentous actinobacteria Frankia is laborious because of the slow growth rate and required biomass needed for these techniques. An efficient and simple colony PCR protocol for Frankia was developed that saved time for analysis of any Frankia strains growing on a plate. Previously, it took 5–6 weeks to get the correct size Frankia colonies on plates and then a minimum of 5 weeks of growth in liquid culture for DNA extraction. With this technique, these colonies could be screened after 5–6 weeks of growth by colony PCR. The procedure used a combination of mechanical and heat treatments and required no added buffers or chemicals. Our results demonstrate rapid and efficient PCR.
KeywordsColony PCR Frankia Genetic tools Molecular technique Mutant identification
CP and VAK designed the study, performed the research, and analysed the data. CP, VAK and LST wrote the paper. All the authors approved the paper.
Partial funding was provided by the New Hampshire Agricultural Experiment Station. This is Scientific Contribution Number 2779. This work was also supported by the USDA National Institute of Food and Agriculture Hatch 022821 (LST), Agriculture and Food Research Initiative Grant 2015-67014-22849 from the USDA National Institute of Food and Agriculture (LST), and the College of Life Science and Agriculture at the University of New Hampshire-Durham.
Compliance with ethical standards
Conflict of interest
The authors declared that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
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