Cycle Sequencing of Filamentous Phage DNA Using a Biotinylated Primer and ΔTaq DNA Polymerase

Abstract

We describe a rapid nonradioactive, double-stranded phage DNA sequencing method using ΔTaq DNA polymerase in cycle reaction for phage peptide-display library screening. This procedure is specific, rapid, sensitive and safe for the sequencing of large numbers of phage peptide-display colonies. In addition, thermal cycle sequencing with this chemiluminescent image detection protocol provides an inexpensive method for completing a large amount of sequencing reactions. Consequently, this method is especially useful for those extensively working on peptide mimics using phage display libraries.