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A mobility-shift detection method for DNA methylation analysis using phosphate-affinity polyacrylamide gel electrophoresis Kinoshita-Kikuta, Emiko Kinoshita-Kikuta Emiko Kinoshita, Eiji Kinoshita Eiji Koike, Tohru Koike Tohru open access Copyright (c) 2008 Elsevier Inc. DNA methylation; Epigenetics Methylation-specific PCR Phosphate-affinity PAGE Phos-tag 490 Herein, we describe a procedure for DNA methylation analysis using the bisulfite-mediated C-to-U conversion of a target DNA followed by methylation-specific polymerase chain reaction (MSP) and phosphate-affinity polyacrylamide gel electrophoresis (PAGE). The MSP was performed using a 1:1 mixture of 5'-phosphorylated methylation-specific and 5'-OH non-methylation-specific primers. The PAGE using an immobilized phosphate-binding tag molecule (i.e., a polyacrylamide-bound dizinc(II) complex, Zn2+–Phos-tag), which selectively captures the 5'-phosphorylated DNA fragment, enabled the mobility-shift detection of the methylation-specific product as a slower migration band. Using this novel procedure, we demonstrated the detection of a methylated cytosine base in a pUC19 plasmid. Academic Press Inc. Elsevier Science 2008-07-01 eng journal article AO https://hiroshima.repo.nii.ac.jp/records/2006206 10.1016/j.ab.2008.03.021 http://dx.doi.org/10.1016/j.ab.2008.03.021 0003-2697 AA00524867 Analytical Biochemistry Analytical Biochemistry 378 1 102 102 104 https://hiroshima.repo.nii.ac.jp/record/2006206/files/AnalBiochem_378_102.pdf 635.0 KB