A mobility-shift detection method for DNA methylation analysis using phosphate-affinity polyacrylamide gel electrophoresis
Analytical Biochemistry Volume 378 Issue 1
Page 102-104
published_at 2008-07-01
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| File | |
| Title ( eng ) |
A mobility-shift detection method for DNA methylation analysis using phosphate-affinity polyacrylamide gel electrophoresis
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| Creator |
Kinoshita-Kikuta Emiko
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| Source Title |
Analytical Biochemistry
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| Volume | 378 |
| Issue | 1 |
| Start Page | 102 |
| End Page | 104 |
| Abstract |
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.
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| Keywords |
DNA methylation; Epigenetics
Methylation-specific PCR
Phosphate-affinity PAGE
Phos-tag
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| NDC |
Medical sciences [ 490 ]
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| Language |
eng
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| Resource Type | journal article |
| Publisher |
Academic Press Inc.
Elsevier Science
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| Date of Issued | 2008-07-01 |
| Rights |
Copyright (c) 2008 Elsevier Inc.
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| Publish Type | Author’s Original |
| Access Rights | open access |
| Source Identifier |
[ISSN] 0003-2697
[DOI] 10.1016/j.ab.2008.03.021
[NCID] AA00524867
[DOI] http://dx.doi.org/10.1016/j.ab.2008.03.021
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