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oai:hiroshima.repo.nii.ac.jp:02006203 2025-02-21T06:56:33Z 1730444907710

A heteroduplex-preferential Tm depressor for the specificity-enhanced DNA polymerase chain reactions Kinoshita, Eiji Kinoshita-Kikuta, Emiko Koike, Tohru Macrocyclic tetraamine Zinc(II) complex Tm depressor Specificity-enhanced PCR 460 A macrocyclic tetraamine zinc(II) complex appended with two quinoline groups, Zn2+.1,7-bis(4-quinolylmethyl)-1,4,7,10-tetraazacyclododecane (Zn2+.Q2-cyclen), was successfully used as a novel additive to suppress non-specific products in DNA polymerase chain reaction (PCR). In the presence of Zn2+.Q2-cyclen, the Tm drop of 20-bp heteroduplexes containing a non-complementary base pair was greater than that of the corresponding homoduplex (i.e., primer DNA). Here, we applied such preferential DNA melting to a specificity-enhanced PCR using micromolar concentrations of Zn2+.Q2-cyclen. We demonstrated the selective amplification of target DNA fragments (i.e., the human heart sodium channel Nav1.5 gene) from genomic DNA or a cDNA library. The optimum condition for the specificity-enhanced PCR could be determined in the concentration range of 1.50 μM of Zn2+.Q2-cyclen. http://purl.org/coar/resource_type/c_6501 Elsevier 2005-02-01 AO 0003-2697 AA00524867 154 Analytical Biochemistry 1 337 160 154 Analytical Biochemistry https://hiroshima.repo.nii.ac.jp/records/2006203 eng 10.1016/j.ab.2004.10.009 15649389 http://dx.doi.org/10.1016/j.ab.2004.10.009 open access Copyright (c) 2004 Elsevier Ltd.