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Abstract
We present a method for the in vitro amplification of > 6.0 kb of DNA flanking a known site. This is accomplished by ligating an oligonucleotide to create an inverted repeat of a portion of the known sequence, followed by single-primer polymerase chain reaction (PCR) amplifications. This method generates a panhandle template following primer extension on the strand of interest. It does not involve template-directed extension from the ligated oligonucleotide, and it is carried out without DNA extractions. We have used this method to amplify 4.5-9.4 kb of DNA flanking the original primer annealing sites directly from human genomic DNA.
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