Evaluation of a Partially-automated Magnetic Bead-based Method for DNA Extraction for Wheat and Barley MAS

Czembor, P.Cz. and Sejbuk, K. and Kleszcz, R. (2014) Evaluation of a Partially-automated Magnetic Bead-based Method for DNA Extraction for Wheat and Barley MAS. Cereal Research Communications, 42 (1). pp. 27-37. ISSN 0133-3720

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Abstract

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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