A two-step electrodialysis method for DNA purification from polluted metallic environmental samples.
Extracting DNA from samples of polluted environments using standard methods often results in low yields of poor-quality material unsuited to subsequent manipulation and analysis by molecular biological techniques. Here, we report a novel two-step electrodialysis-based method for the extraction of DNA from environmental samples. This technique permits the rapid and efficient isolation of high-quality DNA based on its acidic nature, and without the requirement for phenol-chloroform-isoamyl alcohol cleanup and ethanol precipitation steps. Subsequent PCR, endonuclease restriction, and cloning reactions were successfully performed utilizing DNA obtained by electrodialysis, whereas some or all of these techniques failed using DNA extracted with two alternative methods. We also show that his technique is applicable to purify DNA from a range of polluted and nonpolluted samples.
Rodríguez-Mejía
José Luis
JL
Laboratorio de Biología Molecular de Hongos, Centro de Investigación en Biotecnología, Universidad Autónoma del Estado de Morelos, Cuernavaca, Morelos, México.
Martínez-Anaya
Claudia
C
Folch-Mallol
Jorge Luis
JL
Dantán-González
Edgar
E
D002617:Chemical Industry; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification*; D003956:Dialysis / Q000295:instrumentation / Q000379:methods*; D004586:Electrophoresis / Q000379:methods*; D004867:Equipment Design; D006863:Hydrogen-Ion Concentration; D007220:Industrial Waste / Q000032:analysis*; D010084:Oxidation-Reduction; D010578:Petroleum / Q000382:microbiology; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012329:RNA, Bacterial / Q000235:genetics; D012331:RNA, Fungal / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D021521:Ribotyping / Q000379:methods*; D013232:Steel; D014874:Water Pollutants, Chemical / Q000032:analysis*
D004269:DNA, Bacterial; D004271:DNA, Fungal; D007220:Industrial Waste; D010578:Petroleum; D012329:RNA, Bacterial; D012331:RNA, Fungal; D012336:RNA, Ribosomal, 16S; D012337:RNA, Ribosomal, 18S; D014874:Water Pollutants, Chemical; D013232:Steel