Evaluation of selected DNA isolation methods for wheat breeding purposes
Aleksandra Pietrusińska
a.pietrusinska@ihar.edu.plZakład Hodowli i Genetyki Stosowanej, Instytut Hodowli i Aklimatyzacji Roślin w Radzikowie (Poland)
Paweł Cz. Czembor
Zakład Fitopatologii, Instytut Hodowli i Aklimatyzacji Roślin w Radzikowie (Poland)
Abstract
There are many methods of plant DNA isolation. However, most of them are unacceptable for marker assisted breeding since their procedures are long, complex and expensive. Therefore, we evaluated three methods that could be used in marker assisted selection in wheat breeding: DNeasy Plant Mini Kit (Qiagen), DNAzol Direct (Molecular Research Center) and alkaline lysis based on NaOH. All isolated DNA were kept in the same conditions. In order to evaluate stability of DNA matrices, they were tested in PCR reaction after one, ten, thirty and forty days of storage. Each DNA sample was amplified in PCR with twelve SSRs (Simple Sequence Repeats). Three DNA isolation methods were assessed based upon number of the amplified PCR products and their intensity after various periods of DNA storage. Besides the reference method DNeasy Plant Mini Kit, the most stable and acceptable results were obtained with fast DNA extraction method based on NaOH. The last method coupled with microsatellite assisted selection seems to be the most reliable and cost effective in breeding programs, whereas DNAzol Direct method gave results different to that described in the literature.
Keywords:
DNA isolation, wheatReferences
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Authors
Aleksandra Pietrusińskaa.pietrusinska@ihar.edu.pl
Zakład Hodowli i Genetyki Stosowanej, Instytut Hodowli i Aklimatyzacji Roślin w Radzikowie Poland
Authors
Paweł Cz. CzemborZakład Fitopatologii, Instytut Hodowli i Aklimatyzacji Roślin w Radzikowie Poland
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