Effect of divalent metal ions on DNA studied by capillary electrophoresis.
Divalent metal ions, such as Zn(2+), Co(2+), and Ni(2+), are capable of incorporating into DNA under certain conditions to form complexes termed M-DNA. To better understand the effects of these cations on DNA we used capillary electrophoresis (CE). The presence of these metal ions in a typical genotyping buffer led to broad peaks with low fluorescence intensities. In addition, some of the metal-complexed DNA molecules had different electrophoretic mobilities than their normal DNA counterparts. It is likely that the mobility shifts observed in the electropherograms of these affected fragments are due to the divalent cations causing structural changes in the single-stranded DNA. However, as can be seen from the resulting peak shapes, the structure, charge, and/or mass changes due to metal binding are not conserved among all of the DNA fragments. The extent of both peak-broadening and mobility shifts were found to be dependent on the metal cation and its concentration, the length of time that the DNA sample existed in formamide prior to injection into the capillary, and also the fragment size and sequence. These results suggest that the presence of metal ions might be responsible for the poor CE performance that occurs when genotyping certain kinds of DNA samples.
Hartzell
Brittany
B
Department of Chemistry and Biochemistry, Ohio University, Athens, OH 45701, USA.
McCord
Bruce
B
D002413:Cations, Divalent / Q000494:pharmacology*; D002712:Chlorides / Q000494:pharmacology; D003035:Cobalt / Q000494:pharmacology; D004247:DNA / Q000187:drug effects* / Q000302:isolation & purification; D016172:DNA Fingerprinting / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D005296:Ferrous Compounds / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D009532:Nickel / Q000494:pharmacology; D011107:Polylysine / Q000494:pharmacology; D013050:Spectrometry, Fluorescence; D013096:Spermine / Q000494:pharmacology; D017967:Zinc Compounds / Q000494:pharmacology
D002413:Cations, Divalent; D002712:Chlorides; D005296:Ferrous Compounds; D017967:Zinc Compounds; D011107:Polylysine; D013096:Spermine; D003035:Cobalt; C022838:nickel chloride; D009532:Nickel; C016837:zinc chloride; D004247:DNA; C018021:cobaltous chloride; C029451:ferrous chloride