AUTOMATION OF THE MAXAM-GILBERT CHEMICAL SEQUENCING REACTIONS

A practical automated method of Maxam-Gilbert chemical sequencing reac tions that uses solid-phase chromatography methods to purify DNA follo wing chemical modification and cleavage is described in this report. T he automation has primarily been made possible by using specially desi gned BioPak mini-columns, compatible with the Biomek(TM) 1000 automate d workstation, which can be utilized in a manner similar to that of st andard pipet tips. This automated chromatographic sequencing method pr oduces rapid and reliable data as verified by sequencing a known human factor IX exon VIII gene fragment. The procedure presented in this re port is a prototype for a single-fragment reaction and can easily be e xpanded to perform reactions on as many as 8 fragments at a time. The automation eliminates the tedious and time-consuming steps in the orig inal method and increases the rate of sequence acquisition. This techn ology makes the Maxam-Gilbert chemical sequencing protocol more access ible, especially in large-scale, automated sequencing projects.