Plant Molecular Biology Reporter 21: 97, March 2003 9 2003 International Society for Plant Molecular Biology. Printed in Canada. Publish by Abstract
Isolating Conifer DNA: A Superior Polysaccharide Elimination Method TAMSYN M. CROWLEY*, MORLEY S. MURALITHARAN and TREVOR W. STEVENSON School of Biological and Chemical Sciences, Deakin University, Geelong, 3217, Victoria, Australia
Abstract. Genomic DNA was isolated from frozen needles of mature Pinus radiata clones using a modified extraction technique incorporating cetyltrimethylammonium bromide (CTAB) for cell lysis. A high sodium chloride concentration (2 M) was used at 2 stages of the extraction procedure to eradicate polysaccharides, yielding pure genomic DNA suitable for restriction enzyme digestion and PCR amplification. Extractions were scaled down to suit 1.5-mL Eppendorf tubes, allowing easier handling and enhanced sterility. Full textt: This manuscript, in detail, is available only in the electronic version of the Plant Molecular Biology Reporter.
Contents: This article contains Introduction, Materials and Methods, Results and Discussion, 8 references, and 2 illustrations. Illustrations Figure 1. Agarose gel (1.2% w/v) of undigested DNA isolated from mature Pinus radiata needles. Figure 2. Agarose gel (1.5% w/v) of PCR amplification of P radiata DNA using microsatellite primer 857. Key words: conifer, CTAB, genomic DNA, Pinus radiata, polysaccharides
*Author for correspondence, e-mail:
[email protected]; fax: 61 3 5277 2022; ph: 61 3 5277 2848. tEditor's note: Although the scientific content of this manuscript has been reviewed, the full text Web document has not been edited in detail.
