אסיף מאגר המחקר החקלאי

Comparative Analysis of Different DNA Extraction Protocols: A Fast, Universal Maxi-Preparation of High Quality Plant DNA for Genetic Evaluation and Phylogenetic Studies

Year:

1998

Source of publication :

Authors :

Volume :

16

Co-Authors:

Csaikl, U.M., Austrian Research Centre Seibersdorf, Biotechnology Unit, A-2444 Seibersdorf, Austria
Bastian, H., QIAGEN GmbH, Max-Volmer-Strasse 4, D-40724 Hilden, Germany
Brettschneider, R., Ctr. for Appl. Plant Molec. Biology, AMPII, Institute of General Botany, Ohnhorststrasse 18, D-22609 Hamburg, Germany
Gauch, S., QIAGEN GmbH, Max-Volmer-Strasse 4, D-40724 Hilden, Germany
Meir, A., QIAGEN GmbH, Max-Volmer-Strasse 4, D-40724 Hilden, Germany
Schauerte, M., Fed. Res. Ctr. Forest. Forest Prod., Institute for Forest Genetics, Sieker Landstrasse 2, D-22927 Grosshansdorf, Germany
Scholz, F., Fed. Res. Ctr. Forest. Forest Prod., Institute for Forest Genetics, Sieker Landstrasse 2, D-22927 Grosshansdorf, Germany
Sperisen, C., Swiss Fed. Inst. Forest, Snow L., Zürcherstrasse 111, CH-8903 Birmensdorf, Switzerland
Vornam, B., Inst. Forest Genet. and Plant Breed., University of Göttingen, Büsgenweg 2, D-37077 Göttingen, Germany
Ziegenhagen, B., Fed. Res. Ctr. Forest. Forest Prod., Institute for Forest Genetics, Sieker Landstrasse 2, D-22927 Grosshansdorf, Germany

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Abstract:

Four DNA extraction protocols were compared for ability to produce DNA from the leaves or needles of several species: oak, elm, pine, fir, poplar and maize (fresh materials) and rhododendron (silica dried or frozen material). With the exception of maize and poplar, the species are known to be difficult for DNA extraction. Two protocols represented classical procedures for lysis and purification, and the other two were a combination of classical lysis followed by anion exchange chromatography. The DNA obtained from all procedures was quantified and tested by PCR and Southern hybridisation. Test results indicated superiority of one of the four protocols; a combination of CTAB lysis followed by anion exchange chromatography which enabled DNA extraction from all seven species. A second protocol also produced DNA from leaves or needles of all species investigated and was well suited for PCR applications but not Southern hybridisations. The remaining protocols produced DNA from some but not all species tested.

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