Double strand cleavage of genomic DNA at a single site by triple helix formation

Abstract

The sequence-specific cleavage of double helical DNA by restriction endonucleases is essential for many techniques in molecular biology, including gene isolation, DNA sequencing, and recombinant DNA manipulations. With the advent of pulsed-field gel electrophoresis, the separation of large segments of DNA is now possible. However, the recognition sequences of naturally occurring restriction enzymes are in the range of 4-8 base pairs, and hence their sequence specificites may be inadequate for isolating genes from large chromosomes (10^8 base pairs in size) or mapping genomic DNA.