Rapid and direct sequencing of double-stranded DNA using exonuclease III and MALDI-TOF MS.
Application of MALDI-TOF MS to direct sequencing of dsDNA substrates is demonstrated using a strategy that employs exonuclease III digestion of a target sequence. Experimental conditions for exonuclease III have been optimized for this application, including addition of essential divalent metal ion cofactors. A short cation-exchange column was designed to provide efficient sample cleanup and overcome major problems arising from salt interference.
Evans Laboratory of Chemistry, The Ohio State University, Columbus 43210, USA.
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Puapaiboon
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Jai-nhuknan
J A
JA
Cowan
D004247:DNA / Q000032:analysis*; D005090:Exodeoxyribonucleases / Q000737:chemistry*; D009841:Oligonucleotides / Q000737:chemistry; D017422:Sequence Analysis, DNA / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D009841:Oligonucleotides; D004247:DNA; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III