Simple SNP typing assay using a base-discriminating fluorescent probe.
We have developed a new concept involving a single-step homogeneous method for single-nucleotide polymorphism (SNP) typing. In this method, a probe containing base-discriminating fluorescent (BDF) bases is added to a sample solution. BDF base-containing DNA usually shows only a weak fluorescence, but emits a strong blue fluorescence when it recognizes a target base at a specific site in a hybridized strand. By utilizing this feature, a simple mix-and-read SNP typing assay was achieved without any tedious probe-designing or washing processes for exclusion of hybridization error or any addition of DNA-modifying enzymes. This is very different from conventional methods. We simultaneously analyzed a number of samples with ease, with a high accuracy, using our BDF assay.
Okamoto
Akimitsu
A
Department of Synthetic Chemistry and Biological Chemistry, Faculty of Engineering, Kyoto University, Kyoto, 615-8510, Japan. okamoto@sbchem.kyoto-u.ac.jp
Tainaka
Kazuki
K
Ochi
Yuji
Y
Kanatani
Keiichiro
K
Saito
Isao
I
D001483:Base Sequence; D001681:Biological Assay; D004247:DNA / Q000737:chemistry*; D015342:DNA Probes / Q000138:chemical synthesis* / Q000737:chemistry; D005260:Female; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry; D006801:Humans; D008958:Models, Molecular; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D013050:Spectrometry, Fluorescence
D015342:DNA Probes; D005456:Fluorescent Dyes; D004247:DNA