Methods for sequencing large nucleic acid segments and compositions for large sequencing projects

摘要

Methods for rapidly sequencing large nucleic acids by making overlapping fragments and anchored PCR amplification are provided. In the methods, an overlapping set of double stranded nucleic acids is provided, primer binding sites are ligated to the double stranded nucleic acids, and anchored PCR is performed on the resulting primer binding site tagged nucleic acids. PCR amplified fragments are sequenced by chain termination methods. These long distance sequencing methods are used independently, or in conjunction with other sequencing methods, such as shotgun sequencing techniques. Sets of PCR reactions for performing rapid sequencing and kits for practicing the methods are also provided.