The chased polymerase chain reaction (PCR) technique described here is a convenient method enabling the characterization of flanking regions of a known A/T-rich DNA fragment in two different successive steps. The first step includes a modified inverse PCR with inverted tail-to-tail primers, each with 35 overhanging nucleotides for the insertion into a carrier plasmid. The second step consists of a technique similar to a site-directed mutagenesis. The chased PCR technique is simple, quick, versatile and efficient; it improves the inverse PCR technique and may be applied to any ligation-linker method. Consequently, the techniques for PCR-based gene isolation are more suitable for the isolation of missing sequences of A/T-rich or complex DNA.

Journal Section:
Short Communication
Keywords:
A/T-rich and complex DNA, Chased PCR, Glycosylphosphatidylinositol, Inverse PCR, Plasmodium falciparum, apicomplexa, Polymerase chain reaction, Site-directed mutagenesis
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© 2003 S. Karger AG, Basel
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2003
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