Automation of the Maxam-Gilbert chemical sequencing reactions

E. J. Boland, A. Pillai, M. W. Odom, P. Jagadeeswaran

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

A practical automated method of Maxam-Gilbert chemical sequencing reactions that uses solid-phase chromatography methods to purify DNA following chemical modification and cleavage is described in this report. The automation has primarily been made possible by using specially designed BioPak mini-columns, compatible with the Biomek(TM) 1000 automated workstation, which can be utilized in a manner similar to that of standard pipet tips. This automated chromatographic sequencing method produces rapid and reliable data as verified by sequencing a known human factor IX exon VIII gene fragment. The procedure presented in this report is a prototype for a single-fragment reaction and can easily be expanded to perform reactions on as many as 8 fragments at a time. The automation eliminates the tedious and time-consuming steps in the original method and increases the rate of sequence acquisition. This technology makes the Maxam-Gilbert chemical sequencing protocol more accessible, especially in large-scale, automated sequencing projects.

Original languageEnglish (US)
Pages (from-to)1088+1090-1092+1094-1095
JournalBioTechniques
Volume16
Issue number6
StatePublished - Jan 1 1994

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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