Simple SNP typing assay using a base-discriminating fluorescent probe |
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Authors: | Okamoto Akimitsu Tainaka Kazuki Ochi Yuji Kanatani Keiichiro Saito Isao |
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Institution: | Department of Synthetic Chemistry and Biological Chemistry, Faculty of Engineering, Kyoto University, Kyoto, 615-8510, Japan. okamoto@sbchem.kyoto-u.ac.jp |
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Abstract: | 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. |
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