Single base mismatch detection by microsecond voltage pulses.
A single square voltage pulse applied to metal electrodes underneath a silicon dioxide film upon which DNA probes are immobilized allows the discrimination of DNA targets with a single base mismatch during hybridization. Pulse duration, magnitude and slew rate of the voltage pulse are all key factors controlling the rates of electric field assisted hybridization. Although pulses with 1 V, lasting less than 1 ms and with a rise/fall times of 4.5 ns led to maximum hybridization of fully complementary strands, lack of stringency did not allow the discrimination of single base mismatches. However, by choosing pulse conditions that are slightly off the optimum, the selectivity for discriminating single base mismatches could be improved up to a factor approximately 5 when the mismatch was in the middle of the strand and up to approximately 1.5 when the mismatch was on the 5'-end and. These results demonstrate that hybridization with the appropriate electric field pulse provides a new, site-specific, approach to the discrimination of single nucleotide polymorphisms in the sub-millisecond time scale, for addressable DNA microarrays.
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INESC-Microsistemas and Nanotecnologias, INESC MN, Rua Alves Redol, 9, 1000-029 Lisboa, Portugal.
Chu
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Prazeres
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DM
Conde
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JP
D020137:Base Pair Mismatch; D004247:DNA / Q000032:analysis* / Q000737:chemistry* / Q000528:radiation effects; D004252:DNA Mutational Analysis / Q000379:methods*; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods*; D004574:Electromagnetic Fields; D017403:In Situ Hybridization / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D020641:Polymorphism, Single Nucleotide
D004247:DNA