A Non-Radioactive Multiprime Sequencing Method for HIV Genomes.

A manual non-radioactive DNA sequencing protocol was developed for rapid analysis of variable HIV-1 genomes. Sets of up to ten primers were used in one sequencing reaction. After polyacrylamide gel electrophoresis and blotting onto nylon membranes the individual sequences were detected by hybridization with digoxigenin-labelled oligonucleotides and chemiluminescence. The method is applicable to any sequencing project where numerous variants of DNA fragments of several 1000 bp of length are to be analysed.