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Sequencing from Compomers: Using Mass Spectrometry for DNA De-Novo Sequencing of 200+ nt

  • Sebastian Böcker
Part of the Lecture Notes in Computer Science book series (LNCS, volume 2812)

Abstract

One of the main endeavors in today’s Life Science remains the efficient sequencing of long DNA molecules. Today, most de-novo sequencing of DNA is still performed using electrophoresis-based Sanger Sequencing, based on the Sanger concept of 1977. Methods using mass spectrometry to acquire the Sanger Sequencing data are limited by short sequencing lengths of 15–25 nt.

We propose a new method for DNA sequencing using base-specific cleavage and mass spectrometry, that appears to be a promising alternative to classical DNA sequencing approaches. A single stranded DNA or RNA molecule is cleaved by a base-specific (bio-)chemical reaction using, for example, RNAses. The cleavage reaction is modified such that not all, but only a certain percentage of those bases are cleaved. The resulting mixture of fragments is then analyzed using MALDI-TOF mass spectrometry, whereby we acquire the molecular masses of fragments. For every peak in the mass spectrum, we calculate those base compositions that will potentially create a peak of the observed mass and, repeating the cleavage reaction for all four bases, finally try to uniquely reconstruct the underlying sequence from these observed spectra. This leads us to the combinatorial problem of Sequencing From Compomers and, finally, to the graph-theoretical problem of finding a walk in a subgraph of the de Bruijn graph. Application of this method to simulated data indicates that it might be capable of sequencing DNA molecules with 200+ nt.

Keywords

Sample Sequence Mass Spectrometry Data Cleavage Reaction Sequence Candidate Edge Transition 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2003

Authors and Affiliations

  • Sebastian Böcker
    • 1
  1. 1.AG Genominformatik, Technische FakultätUniversität BielefeldBielefeldGermany

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