INFERENCE SEQUENCING BY HYBRIDIZATION

摘要

The inventors present a new strategy for de novo sequencing using oligonucleotide probe arrays which we term Inference Sequencing by Hybridization (ISBH). The ISBH method characterises a target DNA with several small degenerate probe arrays and reconstructs the base sequence of the target via computer algorithm at 100 % accuracy. ISBH returns a target sequence as multiple non-overlapping fragments several hundred to several thousand bases in length. Target DNAs up to 100 kilobases long can potentially be sequenced in a single experiment. A hypothetical ISBH probe array and sequence reconstruction algorithm were designed and tested by computer simulation on 76 DNA sequences obtained from GenBank comprising a total of 2.45 million base pairs. ISBH performs optimally on target DNAs in the range of 15-45 kilobases that have low levels of repeated sequence. We believe that the ISBH approach has great potential to increase the speed of DNA sequencing.