Fluorescent quenching-based quantitative detection of specific DNA/RNA using a BODIPY(R) FL-labeled probe or primer
Author(s) -
Sumiko Kurata
Publication year - 2001
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/29.6.e34
Subject(s) - guanine , oligonucleotide , primer (cosmetics) , biology , dna , cytosine , fluorescence , bodipy , quenching (fluorescence) , microbiology and biotechnology , primer dimer , molecular beacon , biophysics , polymerase chain reaction , biochemistry , nucleotide , gene , multiplex polymerase chain reaction , chemistry , physics , organic chemistry , quantum mechanics
We have developed a simple method for the quantitative detection of specific DNA or RNA molecules based on the finding that BODIPY((R)) FL fluorescence was quenched by its interaction with a uniquely positioned guanine. This approach makes use of an oligonucleotide probe or primer containing a BODIPY((R)) FL-modified cytosine at its 5'-end. When such a probe was hybridized with a target DNA, its fluorescence was quenched by the guanine in the target, complementary to the modified cytosine, and the quench rate was proportional to the amount of target DNA. This widely applicable technique will be used directly with larger samples or in conjunction with the polymerase chain reaction to quantify small DNA samples.
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