Genome walking with 2- to 4-kb steps using panhandle PCR.
Panhandle PCR is a method that permits the rapid retrieval of DNA segments flanking a known DNA sequence. This method extends the application of PCR to the retrieval of DNA where only one end of the DNA sequence is known, so that one can use PCR to walk along an uncharacterized stretch of DNA. In this report, we demonstrate that this method can be applied to the highly specific retrieval of large pieces (2.2-4.4 kb) of human genomic DNA that flank the primer annealing sites.
Jones
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DH
Department of Pediatrics, University of Iowa College of Medicine, Iowa City 52242-1083.
Winistorfer
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SC
D001483:Base Sequence; D016386:Chromosome Walking / Q000379:methods*; D003550:Cystic Fibrosis / Q000235:genetics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA / Q000032:analysis; D004277:DNA, Single-Stranded; D005914:Globins / Q000235:genetics; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*
C505032:CFTR protein, human; D004277:DNA, Single-Stranded; D008565:Membrane Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005914:Globins; D004247:DNA