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dc.contributor.authorAngles d'Auriac, Marc B
dc.date.accessioned2018-08-01T12:47:24Z
dc.date.available2018-08-01T12:47:24Z
dc.date.created2017-02-14T21:57:14Z
dc.date.issued2016
dc.identifier.citationPLoS ONE. 2016, 11 (10), e0165468.nb_NO
dc.identifier.issn1932-6203
dc.identifier.urihttp://hdl.handle.net/11250/2507157
dc.description.abstractAvoiding complementarity between primers when designing a PCR assay constitutes a central rule strongly anchored in the mind of the molecular scientist. 3'-complementarity will extend the primers during PCR elongation using one another as template, consequently disabling further possible involvement in traditional target amplification. However, a 5'-complementarity will leave the primers unchanged during PCR cycles, albeit sequestered to one another, therefore also suppressing target amplification. We show that 5'-complementarity between primers may be exploited in a new PCR method called COMplementary-Primer-Asymmetric (COMPAS)-PCR, using asymmetric primer concentrations to achieve target PCR amplification. Moreover, such a design may paradoxically reduce spurious nontarget amplification by actively sequestering the limiting primer. The general principles were demonstrated using 5S rDNA direct repeats as target sequences to design a speciesspecific assay for identifying Salmo salar and Salmo trutta using almost fully complementary primers overlapping the same target sequence. Specificity was enhanced by using 3'-penultimate point mutations and the assay was further developed to enable identification of S. salar x S. trutta hybrids by High Resolution Melt analysis in a 35 min one-tube assay. This small paradigm shift, using highly complementary primers for PCR, should help develop robust assays that previously would not be considered.nb_NO
dc.language.isoengnb_NO
dc.publisherPublic Library of Sciencenb_NO
dc.rightsNavngivelse 4.0 Internasjonal*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/deed.no*
dc.titleCOMplementary Primer ASymmetric PCR (COMPAS-PCR) Applied to the Identification of Salmo salar, Salmo trutta and Their Hybridsnb_NO
dc.typeJournal articlenb_NO
dc.typePeer reviewednb_NO
dc.description.versionpublishedVersionnb_NO
dc.rights.holder© 2016 Marc B. Anglès d'Auriac.nb_NO
dc.source.pagenumber15nb_NO
dc.source.volume11nb_NO
dc.source.journalPLoS ONEnb_NO
dc.source.issue10nb_NO
dc.identifier.doi10.1371/journal.pone.0165468
dc.identifier.cristin1450594
dc.relation.projectNorsk institutt for vannforskningnb_NO
cristin.unitcode7464,20,15,0
cristin.unitnameAkvakultur
cristin.ispublishedtrue
cristin.fulltextoriginal
cristin.qualitycode1


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