Molecular markers
DNA sequencing
Real time single molecule DNA sequence determination




Topic Completed: 1 February 2011

Revised: 15 February 2019

Copyright: 2008-2018, PathologyOutlines.com, Inc.

PubMed Search: Real time single molecule DNA sequence[TIAB] determination



Rodney E. Shackelford, D.O., Ph.D.
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Table of Contents
Definition / general
Cite this page: Shackelford R. Real time single molecule DNA sequence determination. PathologyOutlines.com website. http://www.pathologyoutlines.com/topic/molecularpathdnaseqrealtime.html. Accessed June 19th, 2019.
Definition / general
  • VisiGen Biotechnologies has engineered a DNA polymerase which acts as a real time sensor of modified nucleotides
  • This method uses DNA synthesis, employing a DNA polymerase with a donor fluorescent dye close to the polymerase active site where nucleotides to be incorporated are selected
  • The four dNTPs are also modified, with each having a different acceptor dye compatible with the polymerase donor dye
  • At dNTP incorporation, the donor dye transfers fluorescent resonant energy to the acceptor dye, resulting in measurable light release
  • The four different acceptor dyes give off different wave length emissions, allowing the specific dNTP incorporated to be determined
  • The advantage of this system is that it can sequence DNA at the same rate at the polymerase incorporates dNTPs - several hundred moieties per second, with read lengths of around 1 kb, longer than possible with any current platform
  • This technology may eventually allow incredible sequencing feats, such a sequencing the entire human genome in one day for ~$1,000 (N Biotechnol 2009;25:195)
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