New Methods for the Solid-Phase Sequence Analysis of Nucleic Acid Fragments Using the Sanger Dideoxy Procedure

3′-Terminal tailing of a given DNA fragment with 5-bromodeoxyuridine-5 triphosphate or biotinylated deoxyuridine-5′-triphosphate and deoxynucleotidyl terminal transferase allows its immobilization to an anti-bromo-deoxyuridine-antibody column or to a streptavidin column. The immobilized DNA could be subjected to enzymatic sequencing following the usual protocol. The dideoxy-nucleotide-terminated fragments were eluted with buffer containing formamide / dye mixture and directly applied to gel electrophoresis, allowing reading of ca. 600 bases. Several "sequencing cycles" could be performed with the same DNA column. Semi-mechanization of the process is described.