Comparative Analysis of Different DNA Extraction Protocols: A Fast, Universal Maxi-Preparation of High Quality Plant DNA for Genetic Evaluation and Phylogenetic Studies
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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.
Abbreviations: CTAB, hexadecyltrimethylammonium bromide; EtOH, Ethanol; TBE, tris-borate-EDTA.
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- Carlson JE, Tulsieram LK, Glaubitz JC, Luk VWK, Kauffeldt C and Rutledge R (1991) Segregation of random amplified DNA markers in F1 progeny of conifers. Theor Appl Genet 83: 194-200.Google Scholar
- Degen B, Ziegenhagen B, Gillet E and Scholz F (1995) Computer-Aided Search for Codominant Markers in Complex Haploid DNA Banding Patterns - A Case Study in Abies albaMILL. Silvae Genetica 44: 274–282.Google Scholar
- Dellaporta SL, Wood J and Hicks JB (1983) A plant DNA minipreparation: Version II. Plant Mol Biol Rep 1: 19-21.Google Scholar
- Demesure B, Sodzi N and Petit RJ (1995) A set of universal primers for amplification of polymorphic non-coding regions of mitochondrial and chloroplast DNA in plants. Mol Ecol 4: 129-131.Google Scholar
- Doyle JJ and Doyle JL (1987) A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull 19: 11-15.Google Scholar
- Doyle JJ and Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12: 13-15.Google Scholar
- Feinberg AP and Vogelstein B (1984) A technique for radiolabelling DNA restriction fragments to high specific activity. Analyt Biochem 137: 266-269.Google Scholar
- Gerlach WL and Bedbrook JR (1979) Cloning and characterization of ribosomal RNA genes from wheat and barley. Nucleic Acids Res 7: 1869-1885.Google Scholar
- Guillemaut P and Maréchal-Drouard L (1992) Isolation of plant DNA: A fast, inexpensive and reliable method. Plant Mol Biol Rep 10: 60-65.Google Scholar
- Pandey RN, Adams RP and Flournoy LE (1996) Inhibition of random amplified polymorphic DNAs (RAPDs) by plant polysaccharides. Plant Mol Biol Rep 14(1): 17-22.Google Scholar
- Ziegenhagen B, Guillemaut P and Scholz F (1993) A procedure for mini-preparations of genomic DNA from needles of silver fir (Abies albaMill.). Plant Mol Biol Rep 11: 117-121.Google Scholar
- Ziegenhagen B and Fladung M (1997) Variation in psbCgene region of gymnosperms and angiosperms as detected by a single restriction site polymorphism. Theor Appl Genet 94: 1065-1071.Google Scholar