期刊名称:Proceedings of the National Academy of Sciences
印刷版ISSN:0027-8424
电子版ISSN:1091-6490
出版年度:1989
卷号:86
期号:22
页码:8902-8906
DOI:10.1073/pnas.86.22.8902
语种:English
出版社:The National Academy of Sciences of the United States of America
摘要:We describe a method for characterizing DNA segments that combines limited sequencing with size separation of restriction fragments. As part of a multistep procedure, 5' overhangs of unknown sequence are generated by cleavage with a class IIS restriction enzyme. After labeling of these ends by using dideoxynucleotides tagged with distinctive fluorescent dyes, the restriction fragments are analyzed by polyacrylamide gel electrophoresis and detection of fluorescent emissions using a commercially available DNA sequencer. The nucleotide-specific fluorescent signatures permit determination of the terminal sequence for each labeled end. The set of labeled fragments, characterized by both size and terminal sequence, constitutes a fingerprint that can be used to compare DNA segments for overlap or relatedness. The inclusion of terminal sequence data dramatically increases the information content of the fingerprint, making comparisons more reliable and efficient than those based upon size alone.