A robust and cost-effective method for DNA isolation from satureja species (lamiaceae)
Apstrakt
Aromatic species of the genus Satureja are rich in secondary metabolites that interfere with DNA isolation procedures. 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 procedures (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 mu g 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 i...s suitable for the isolation of high quantities of pure DNA from Satureja spp.
Ključne reči:
Satureja / polyvinylpyrrolidone 10 / polysaccharides / polyphenols / high salt concentration / DNA isolation protocol / activated charcoalIzvor:
Archives of Biological Sciences, 2014, 66, 1, 285-297Izdavač:
- Srpsko biološko društvo, Beograd, i dr.
Finansiranje / projekti:
- Mikromorfološka, fitohemijska i molekularna istraživanja biljaka - sistematski, ekološki i primenljivi aspekti (RS-MESTD-Basic Research (BR or ON)-173029)
- Molekularni mehanizmi odgovora biljaka na abiotički stres-uloga transkripcionih faktora i malih RNK i analiza genetičkog diverziteta biljnih kultura od interesa za poljoprivredu i biotehnologiju (RS-MESTD-Basic Research (BR or ON)-173005)
DOI: 10.2298/ABS1401285D
ISSN: 0354-4664
WoS: 000330552400037
Scopus: 2-s2.0-84892403946
Institucija/grupa
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Dodos, Tanja AU - Aleksić, Jelena M. AU - Rajcević, N. AU - Marin, P. D. PY - 2014 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/766 AB - Aromatic species of the genus Satureja are rich in secondary metabolites that interfere with DNA isolation procedures. 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 procedures (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 mu g 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. PB - Srpsko biološko društvo, Beograd, i dr. T2 - Archives of Biological Sciences T1 - A robust and cost-effective method for DNA isolation from satureja species (lamiaceae) EP - 297 IS - 1 SP - 285 VL - 66 DO - 10.2298/ABS1401285D ER -
@article{ author = "Dodos, Tanja and Aleksić, Jelena M. and Rajcević, N. and Marin, P. D.", year = "2014", abstract = "Aromatic species of the genus Satureja are rich in secondary metabolites that interfere with DNA isolation procedures. 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 procedures (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 mu g 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.", publisher = "Srpsko biološko društvo, Beograd, i dr.", journal = "Archives of Biological Sciences", title = "A robust and cost-effective method for DNA isolation from satureja species (lamiaceae)", pages = "297-285", number = "1", volume = "66", doi = "10.2298/ABS1401285D" }
Dodos, T., Aleksić, J. M., Rajcević, N.,& Marin, P. D.. (2014). A robust and cost-effective method for DNA isolation from satureja species (lamiaceae). in Archives of Biological Sciences Srpsko biološko društvo, Beograd, i dr.., 66(1), 285-297. https://doi.org/10.2298/ABS1401285D
Dodos T, Aleksić JM, Rajcević N, Marin PD. A robust and cost-effective method for DNA isolation from satureja species (lamiaceae). in Archives of Biological Sciences. 2014;66(1):285-297. doi:10.2298/ABS1401285D .
Dodos, Tanja, Aleksić, Jelena M., Rajcević, N., Marin, P. D., "A robust and cost-effective method for DNA isolation from satureja species (lamiaceae)" in Archives of Biological Sciences, 66, no. 1 (2014):285-297, https://doi.org/10.2298/ABS1401285D . .