Test of the potential of a dATP surrogate for sequencing via MALDI-MS.

TitleTest of the potential of a dATP surrogate for sequencing via MALDI-MS.
Publication TypeJournal Article
Year of Publication1997
AuthorsJacutin, S, Zhang, AJ, Russell, DH, Gibbs, RA, Burgess, K
JournalNucleic Acids Res
Date Published1997 Dec 15
KeywordsDeoxyadenine Nucleotides, DNA Polymerase I, DNA-Directed DNA Polymerase, Mass Spectrometry, Nucleotides, Pyrroles, Retroviridae, RNA-Directed DNA Polymerase, Sequence Analysis, DNA, Substrate Specificity, Taq Polymerase, Templates, Genetic

1-(2'-Deoxy-beta-d-ribofuranosyl)-3-nitropyrrole phosphate was incorporated into a DNA decamer and analyzed via matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). The extent and composition of the various fragment peaks were compared with those in the MALDI-MS spectrum of dT4AT5. The nitropyrrole-containing oligomer proved to be more robust. Two different DNA template assays were then used to attempt to identify DNA replicating enzymes that would incorporate the corresponding triphosphate, i.e. 1-(2'-deoxy-beta-d-ribofuranosyl)-3-nitropyrrole triphosphate (dXTP). It was shown that dXTP was not incorporated by some enzymes and it inhibited others. However, DNA polymerase I Klenow fragment and avian myeloblastosis virus reverse transcriptase incorporated dXTP in place of dATP and then replicated the template overhang in the usual way. The potential of dXTP as a surrogate for dATP in DNA sequencing with MALDI-MS analysis is discussed.

Alternate JournalNucleic Acids Res
PubMed ID9396818
PubMed Central IDPMC147155

Similar Publications