Rapid and Direct Sequencing of Double-Stranded DNA Using Exonuclease III and MALDI-TOF MS
| Autor: | Jaran Jai-Nhuknan, James A. Cowan, Uraiwan Puapaiboon |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
Exonuclease III
Chromatography biology Direct sequencing Oligonucleotides Nucleic acid sequence DNA Sequence Analysis DNA Mass spectrometry Molecular biology Analytical Chemistry Matrix-assisted laser desorption/ionization chemistry.chemical_compound Exodeoxyribonucleases chemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Divalent metal ions biology.protein Double stranded |
| Zdroj: | Analytical Chemistry. 72:3338-3341 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/ac000181n |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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