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Method for sequencing synthetic oligodeoxynucleotide phosphorothioates

J Tang1, A M Roskey, S Agrawal

  • 1Hybridon, Inc., Worcester, Massachusetts 01605.

Analytical Biochemistry
|July 1, 1993
PubMed
Summary
This summary is machine-generated.

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A modified Sanger sequencing method enables the analysis of oligodeoxynucleotide phosphorothioates. This technique involves ligating the target molecule to a helper oligonucleotide for primer extension and subsequent gel electrophoresis analysis.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Oligodeoxynucleotide phosphorothioates are synthetic nucleic acid analogs with therapeutic potential.
  • Accurate sequencing of these modified oligonucleotides is crucial for quality control and research.
  • Existing sequencing methods may not be optimal for phosphorothioate backbone modifications.

Purpose of the Study:

  • To describe a modified Sanger sequencing method for oligodeoxynucleotide phosphorothioates.
  • To provide a reliable technique for analyzing the sequence of these modified nucleic acids.

Main Methods:

  • The method involves ligating the synthetic oligodeoxynucleotide phosphorothioate to a helper oligonucleotide.
  • The helper oligonucleotide contains a region complementary to the T7 primer.

Related Experiment Videos

  • DNA polymerase extends a 5'-labeled T7 primer onto the ligated construct.
  • The extended product is analyzed using gel electrophoresis.

    • Main Results:

    • The described method allows for the sequencing of oligodeoxynucleotide phosphorothioates.
    • Successful primer extension and subsequent analysis on gel electrophoresis were demonstrated.

    Conclusions:

    • This modified Sanger sequencing approach provides a viable method for determining the sequence of oligodeoxynucleotide phosphorothioates.
    • The technique facilitates the characterization of synthetic oligonucleotides with modified backbones.