Detection of simple mutations and polymorphisms in large genomic regions

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Show simple item record Sokurenko, E Tchesnokova, V Yeung, A Oleykowski, CA Trintchina, E Hughes, KT Rashid, RA Brint, MR Moseley, S Lorey, S 2009-12-21T02:32:59Z 2001-01
dc.identifier.citation Nucleic Acids Research, 2001, 29 (22), pp. 1 - 8
dc.identifier.issn 1362-4962
dc.identifier.other C1 en_US
dc.description.abstract We have developed a novel technology that makes it possible to detect simple nucleotide polymorphisms directly within a sample of total genomic DNA. It allows, in a single Southern blot experiment, the determination of sequence identity of genomic regions with a combined length of hundreds of kilobases. This technology does not require PCR amplification of the target DNA regions, but exploits preparative size-fractionation of restriction-digested genomic DNA and a newly discovered property of the mismatch-specific endonuclease CEL I to cleave heteroduplex DNA with a very high specificity and sensitivity. We have used this technique to detect various simple mutations directly in the genomic DNA of isogenic pairs of recombinant Pseudomonas aeruginosa, Escherichia coli and Salmonellaisolates. Also, by using a cosmid DNA library and genomic fractions as hybridization probes, we have compared total genomic DNA of two clinical P.aeruginosa clones isolated from the same patient, but exhibiting divergent phenotypes. The mutation scan correctly detected a GA insertion in the quorum-sensing regulator gene rhlR and, in addition, identified a novel intragenomic polymorphism in rrn operons, indicating very high stability of the bacterial genomes under natural non-mutator conditions.
dc.format Y
dc.publisher Oxford University Press
dc.title Detection of simple mutations and polymorphisms in large genomic regions
dc.type Journal Article
dc.parent Nucleic Acids Research
dc.journal.volume 22
dc.journal.volume 29
dc.journal.number 22 en_US
dc.publocation London, UK en_US
dc.identifier.startpage 210 en_US
dc.identifier.endpage 213 en_US IIS en_US
dc.personcode 100098
pubs.embargo.period Not known
pubs.organisational-group /University of Technology Sydney
pubs.organisational-group /University of Technology Sydney/Faculty of Science
utslib.copyright.status Closed Access 2015-04-15 12:17:09.805752+10
utslib.collection.history Closed (ID: 3)

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