Direct exponential amplification and sequencing (DEXAS) of genomic DNA.
In order to supply a sufficient amount of template molecules for DNA sequence determination, cloning into plasmids and subsequent plasmid purification, 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 genomic DNA and thus eliminates the need for template amplification and preparation. It relies on the simultaneous amplification of a target sequence 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.
Kilger
C
C
Institute of Zoology, University of Munich, Germany.
Pääbo
S
S
D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000235:genetics*; D004272:DNA, Mitochondrial / Q000032:analysis / Q000235:genetics; D005240:Feasibility Studies; D005784:Gene Amplification; D015894:Genome, Human; D006801:Humans; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000379:methods*; D013698:Templates, Genetic
D004272:DNA, Mitochondrial; D004247:DNA