A partially automated method of DNA isolation (a Freedom Evo workstation coupled with a paramagnetic bead-based DNA isolation kit from Machery-Nagel) was evaluated in terms of DNA quantity, purity, and quality in comparison with a conventional manual method (the membrane-based Qiagen Plant DNeasy kit). In addition, the efficiency in terms of time and cost was evaluated. The partially automated method enabled the isolation from each sample of DNA of good A260/A280 ratio and sufficient quantity to serve as a template for dozens of PCR amplifications suitable for conventional marker-assisted selection applications. However, lower PCR efficiencies, presumably due to lower DNAquality, were obtained with DNA extracted by the partially automated method as compared with the Qiagen method. The partially automated method presented here was shown to be superior to the conventional Qiagen method in that the costs and time required were reduced by ca. 50%. The Freedom Evo workstation coupled with Machery-Nagel DNA isolation kit appeared advantageous for high-throughput applications required for plant molecular breeding.
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Rajora OPProtocol: A high-throughput DNA extraction system suitable for conifersPlant Methods2008420)| false
Dayteg, C., Tuvesson, S., Merker, A., Jahoor, A., Kolodinska-Brantestam, A. 2007. Automation of DNA marker analysis for molecular breeding in crops: practical experience of a plant breeding company. Plant Breeding 126:410–415.
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Kolodinska-Brantestam AAutomation of DNA marker analysis for molecular breeding in crops: practical experience of a plant breeding companyPlant Breeding2007126410415)| false
Macherey-Nagel 2012. Genomic DNA from plant. User manual NucleoMag 96 Plant, February 2012/Rev.02.
Macherey-Nagel , '', in User manual NucleoMag 96 Plant, February 2012/Rev.02, (2012) -.
Macherey-Nagel User manual NucleoMag 96 Plant, February 2012/Rev.022012)| false
Peist, R., Honsel, D., Twieling, G., Löffert, D. 2001. PCR inhibitors in plant DNA preparations. QIAGEN News 3:7–9.
Löffert D, 'PCR inhibitors in plant DNA preparations' (2001) 3QIAGEN News: 7-9.
Löffert DPCR inhibitors in plant DNA preparationsQIAGEN News2001379)| false
Ramsay, L., Macaulay, M., Ivanissevich, S., MacLean, K., Cardle, L., Fuller, J., Edwards, K.J., Tuvesson, S., Morgante, M., Massari, A., Maestri, E., Marmiroli, N., Sjakste, T., Ganal, M., Powell, W., Waugh, R. 2000. A simple sequence repeat-based linkage map of barley. Genetics 156:1997–2005.
Waugh R, 'A simple sequence repeat-based linkage map of barley' (2000) 156Genetics: 1997-2005.
Waugh RA simple sequence repeat-based linkage map of barleyGenetics200015619972005)| false