Open AccessLabel-free hybridization detection of a single nucleotide mismatch by immobilization of molecular beacons on an agarose film
Author(s) -
Hong Wang,
Jiong Li,
Heping Li,
Quanjun Liu,
Qian Mei,
Yijin Wang,
Jijun Zhu,
gyue He,
Lu Zhang
Publication year - 2002
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/gnf061
Subject(s) - molecular beacon , agarose , biology , molecular probe , dna , microbiology and biotechnology , nucleic acid thermodynamics , beacon , computational biology , genetics , oligonucleotide , base sequence , computer science , real time computing
We developed a new technique to immobilize a set of molecular beacons on an agarose film-coated slide and found that it has the ability to identify a single nucleotide difference in label-free DNA targets. The annealing properties, specificity and hybridization dynamics of the present technique were compared with those of the conventional technique that directly immobilizes molecular beacons on a planar glass slide. It is demonstrated that the molecular beacon array on an agarose film has high quench efficiency, an excellent discrimination ratio for single nucleotide mismatches and a short detection time. We hypothesize that such a low fluorescence background and high specificity molecular beacon array will find practical applications in label-free, high-throughput mutation analysis and disease diagnosis.