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
アクセス数 : 708 件
ダウンロード数 : 228 件
今月のアクセス数 : 1 件
今月のダウンロード数 : 2 件
この文献の参照には次のURLをご利用ください : https://ir.lib.hiroshima-u.ac.jp/00025928
| File | |
| Title ( eng ) |
A mobility-shift detection method for DNA methylation analysis using phosphate-affinity polyacrylamide gel electrophoresis
|
| Creator |
Kinoshita-Kikuta Emiko
|
| Source Title |
Analytical Biochemistry
|
| 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.
|
| Keywords |
DNA methylation; Epigenetics
Methylation-specific PCR
Phosphate-affinity PAGE
Phos-tag
|
| NDC |
Medical sciences [ 490 ]
|
| Language |
eng
|
| Resource Type | journal article |
| Publisher |
Academic Press Inc.
Elsevier Science
|
| Date of Issued | 2008-07-01 |
| Rights |
Copyright (c) 2008 Elsevier Inc.
|
| 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
|
