Significant enhancement of fluorescence on hybridization of a molecular beacon to a target DNA in the presence of a site-specific DNA nickase.
We have developed a simple isothermal (55 degrees C) reaction that permits detection of DNA targets using only two components: a molecular beacon and a site-specific DNA nickase without deoxyribonucleotide triphosphates and primers. The loop sequence of the molecular beacon should contain a DNA nickase recognition site. The nickase-molecular beacon (NMB) combination permits a 100-fold increase in fluorescent signal. The applications of the NMB assay for enhancement of fluorescent signal in some isothermal methods are discussed.
Zheleznaya
Ludmila A
LA
Institute of Theoretical and Experimental Biophysics, 142290 Poushchino, Russian Federation.
Kopein
Damir S
DS
Rogulin
Evgeniy A
EA
Gubanov
Sergey I
SI
Matvienko
Nikolay I
NI
D004247:DNA / Q000737:chemistry*; D015342:DNA Probes / Q000737:chemistry*; D003850:Deoxyribonuclease I / Q000737:chemistry*; D005453:Fluorescence; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013696:Temperature; D013997:Time Factors
D015342:DNA Probes; D009838:Oligodeoxyribonucleotides; D004247:DNA; D003850:Deoxyribonuclease I