Analysis of Nucleotide Sequence Variations by Solid-Phase Minisequencing

  • Anu Suomalainen
  • Ann-Christine Syvänen
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 65)

Abstract

The Sanger dideoxy-nucleotide sequencing method has been simplified by a number of methodological improvements, such as the use of the PCR technique for generating DNA templates in sufficient quantities for sequencing, the use of affinity-capture techniques for convenient and efficient purification of the PCR fragments for sequencing, the development of laboratory robots for carrying out the sequencing reactions, and the development of instruments for automatic on-line analysis of fluorescent products of the sequencing reactions. Despite these technical improvements, the requirement for gel electrophoretic separation remains an obstacle, when sequence analysis of large numbers of samples are needed, as in DNA diagnosis, or in the analysis of sequence variation for genetic, evolutionary, or epidemiological studies.

Keywords

EDTA Agarose Bromide Electrophoresis MgCl2 

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Copyright information

© Humana Press Inc., Totowa, NJ 1996

Authors and Affiliations

  • Anu Suomalainen
    • 1
  • Ann-Christine Syvänen
    • 2
  1. 1.Department of Human Molecular GeneticsNational Public Health InstituteHelsinkiFinland
  2. 2.Department of Biochemistry and BiotechnologyRoyal Institute of TechnologyStockholmSweden

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