A hybrid-specific, polymerase chain reaction-based amplification approach for chromosomal walking.
Here we report the development of an accurate and efficient genome walking approach based on ligation-mediated polymerase chain reaction (PCR) and magnetic capture hybrid selection technique. This approach overcomes the nonspecific amplification products that often occur in similar PCR-based methods. Our strategy was successfully applied for the cloning of the promoter region of the Cc RNase gene. This rapid, cost-effective, and sensitive protocol can easily be extended for use in the isolation and amplification of any target sequences for which only partial information is known such as identification of the position of transposable elements and integrated viral DNA sequences.
Rampias
Theodoros N
TN
Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Athens, Panepistimioupolis, Athens, Greece.
Fragoulis
Emmanuel G
EG
Sideris
Diamantis C
DC
D000818:Animals; D033622:Ceratitis capitata / Q000235:genetics; D016386:Chromosome Walking / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*