Application Guide - QIAxcel Advanced

84 QIAxcel Advanced Application Guide 10/2016 Samples were analyzed with the QIAxcel Advanced System and QIAxcel DNA High Resolution Kit using method OM1200 with an additional 120-second separation time. QX Alignment Marker 50 bp/5 kb was included in the run. QIAxcel ScreenGel software v1.0.1.0 was used to run the samples and export the raw .xml or .csv data. The QIAxcel plugin v1.00 was used to import the raw fingerprint files to BioNumerics version 6.6, using an OD range of 20,000 points and a normalized track resolution of 2500 points as the fingerprint conversion setting. Results PCR-amplified ISSR sequences of individual C. michiganensis subsp. michiganensis strains were resolved, showing well separated, sharp bands in the range of 500–3000 bp. Figure 1 is a dendrogram displaying the relatedness of the isolated strains. Conclusions • The QIAxcel Advanced System gave high-resolution electropherograms, with high bootstrap values, objective band scoring, and minimized background variation. It enables high- throughput analyses of samples in the 96-well plate format. • The analysis data stored in the QIAxcel ScreenGel software can easily be transferred to BioNumerics via the QIAxcel plugin. This enables rapid, reproducible, inexpensive genotyping. Furthermore, it contains integrated analysis and visualization tools for data mining, clustering, identification and statistical analyses. This functionality enables decisive conclusions to be reached. • The combination of the QIAxcel Advanced System and BioNumerics should prove a useful approach in isolating sources of infectious plant diseases, based on the results of this study using Clavibacter michiganesis subsp. michiganesis as a model. References 1. de León, L., Siverio, F., López, M.M., and Rodríguez, A (2011) Clavibacter michiganensis subsp. michiganensis, a seed-borne tomato pathogen: healthy seeds are still the goal. Plant Disease 95, 1328. 2. Anonymous (2005) Clavibacter michiganensis subsp. michiganensis. EPPO Bulletin 35, 275. 3. Strider D.L. (1969) Bacterial canker of tomato caused by Corynebacterium michiganense: a literature review and bibliography: North Carolina Agricultural Experiment Station 4. Ammiraju, J.S.S. et al. (2001) Identification of Inter Simple Sequence Repeat (ISSR) markers associated with seed size in wheat. Theor. Appl. Genet. 102 726. 5. Baysal, Ö. et al. (2010) Clavibacter michiganensis subsp. michiganensis: tracking strains using their genetic differentiations by ISSR markers in Southern Turkey. Physiol. and Mol. Plant Pathol. 75, 113 6. Pitcher, D.G., Saunders, N.A., and Owen, R.J. (1989) Rapid extraction of bacterial genomic DNA with guanidium thiocyanate. Lett. in Appl. Microbiol. 8, 151 in wheat. Theor. Appl. Genet. 102726.

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