A robust and cost-effective DNA isolation method for Satureja species (Lamiaceae)
2014
Dodoš, Tanja (Faculty of Biology, Belgrade (Serbia). Institute of Botany and Botanical Garden Jevremovac) | Rajčević, Nemanja (Faculty of Biology, Belgrade (Serbia). Institute of Botany and Botanical Garden Jevremovac) | Aleksić, Jelena (Institute of Molecular Genetics and Genetic Engineering (IMGGE), Belgrade (Serbia). Laboratory for Plant Molecular Biology) | Marin, Petar D. (Faculty of Biology, Belgrade (Serbia). Institute of Botany and Botanical Garden Jevremovac)
Aromatic species of the genus Satureja are rich in secondary metabolites that interfere with DNA isolation pro- cedures. Four protocols based on the standard CTAB DNA extraction protocol of Doyle and Doyle (1987) were tested in six savory taxa. The polyphenol adsorbents activated charcoal and/or polyvinylpyrrolidone 10 were employed in three pro- cedures (B, C and D); for the elimination of polysaccharides, 4M NaCl was applied in the latter two. The highest DNA yield was obtained with Protocol D and averaged 1420.7±398.3 μg DNA/g of dry leaf tissue. Optimal values of the absorbance ratio 260/280 of all DNA solutions revealed the absence or only negligible contamination by proteins. Contamination by polysaccharides inferred from the absorbance ratio 260/230 showed that Protocol C provided the least contaminated material (average of 1.7±0.4). Enzymatic reactions of DNA solutions obtained by Protocol D showed amplification of both loci in all individuals. In conclusion, Protocol D is suitable for the isolation of high quantities of pure DNA from Satureja spp.
显示更多 [+] 显示较少 [-]