DIRECT EXPONENTIAL AMPLIFICATION AND SEQUENCING (DEXAS) OF GENOMIC DNA

In order to supply a sufficient amount of template molecules for DNA s equence determination, cloning into plasmids and subsequent plasmid pu rification, or amplification via the PCR, are generally used. Here, we present a method - 'direct exponential amplification and sequencing' or 'DEXAS' - that permits direct sequence determination from whole gen omic DNA and thus eliminates the need for template amplification and p reparation. It relies on the simultaneous amplification of a target se quence and the determination of its sequence using dideoxyterminators in a two-step cycling reaction. DEXAS can be applied to single as well as multi-copy genomic sequences, and can easily be automated.