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  • Title: A mobility shift detection method for DNA methylation analysis using phosphate affinity polyacrylamide gel electrophoresis.
    Author: Kinoshita-Kikuta E, Kinoshita E, Koike T.
    Journal: Anal Biochem; 2008 Jul 01; 378(1):102-4. PubMed ID: 18394999.
    Abstract:
    We describe a procedure for DNA methylation analysis using the bisulfite-mediated cytosine-to-uracil 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, Zn(2+)-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.
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