Analysis of enzymatic DNA sequencing reactions by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry
- 1 January 1995
- journal article
- research article
- Published by Wiley in Rapid Communications in Mass Spectrometry
- Vol. 9 (10) , 942-947
- https://doi.org/10.1002/rcm.1290091015
Abstract
The products from base-specific, dideoxy-nucleotide chain-termination DNA sequencing reactions catalyzed by the modified T7 DNA polymerase have been analyzed by using the technique of matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometry. Preliminary experiments were performed to determine detection limits for a synthetic mixture of mixed-base single-stranded DNA which contained a 14-mer, a 21-mer, and a 41-mer; acceptable spectra, showing peaks for each component, were obtainable for samples that contained as little as 5 fmol per component. Initial sequencing reactions were therefore carried out on 2-pmol amounts of a short synthetic template that was 45 nucleotides in length, employing 2 pmol of 12-mer as the primer strand. This provided readable sequence information out to the 19th base past the primer. Using a 21-mer primer, nearly the entire sequence of the template could be read.Keywords
This publication has 11 references indexed in Scilit:
- Mass Resolution Improvement by Incorporation of Pulsed Ion Extraction in a Matrix-Assisted Laser Desorption/Ionization Linear Time-of-Flight Mass SpectrometerAnalytical Chemistry, 1995
- Picolinic acid as a matrix for laser mass spectrometry of nucleic acids and proteinsRapid Communications in Mass Spectrometry, 1994
- Detection of 500‐nucleotide DNA by laser desorption mass spectrometryRapid Communications in Mass Spectrometry, 1994
- Time-of-Flight Mass Spectrometry Of Underivatized Single-Stranded DNA Oligomers by Matrix-Assisted Laser DesorptionAnalytical Chemistry, 1994
- The analysis of mock DNA sequencing reactions using matrix‐assisted laser desorption/ionization mass spectrometryRapid Communications in Mass Spectrometry, 1993
- Matrix‐assisted laser desorption time‐of‐flight mass spectrometry of oligonucleotides using 3‐hydroxypicolinic acid as an ultraviolet‐sensitive matrixRapid Communications in Mass Spectrometry, 1993
- Ion stability of nucleic acids in infrared matrix-assisted laser desorption/ionization mass spectrometryNucleic Acids Research, 1993
- [8] Direct sequencing of polymerase chain reaction productsPublished by Elsevier ,1993
- Effect of manganese ions on the incorporation of dideoxynucleotides by bacteriophage T7 DNA polymerase and Escherichia coli DNA polymerase I.Proceedings of the National Academy of Sciences, 1989
- DNA sequencing with chain-terminating inhibitorsProceedings of the National Academy of Sciences, 1977