2010 Fiscal Year Final Research Report
Development of DNA methylation measurement device using nano-carbon film electrode
| Project/Area Number |
20350042
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Analytical chemistry
|
|---|
| Research Institution | National Institute of Advanced Industrial Science and Technology |
|---|
Principal Investigator |
NIWA Osamu National Institute of Advanced Industrial Science and Technology, バイオメディカル研究部門, 副研究部門長 (70392644)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
UMEMURA Shigeru 千葉工業大学, 工学部, 教授 (70316800)
SUZUKI Koji 慶應義塾大学, 理工学部, 教授 (80154540)
|
|---|
| Project Period (FY) |
2008 – 2010
|
| Keywords | メチル化DNA / チップ分析 / 電気化学検出 / センサ |
|---|
| Research Abstract |
We developed direct electrochemical detection of DNA methylation using nanocarbon film formed by electron cyclotron resonance(ECR) sputtering. The developed method is based on oxidation potential difference between cytosine and methylated cytosine without any labeling. By optimizing electrode surface using electro- chemical pretreatment, we used this film to quantitatively detect single cytosine methylation regardless of the methylation position in the sequence including retino- blastoma gene fragment (24 mer). The nanocarbon film electrode also allowed us to realize the detection of DNA methylation ratios by measuring methylated 5'-cytosine- phosphoguanosine (CpG) repetition oligonucleotides (60 mer) with different methylation ratio. To improve the sensitivity for detecting the bases in the CpG oligonucleotides, we successfully used an exonuclease P1 to digest the target CpG oligonucleotide to an identical mononucleotide 5'-NMP. To develop the DNA methylation detection device, the microdisk array electrode which further improves sensitivity was fabricated using nanocarbon film which suppress biomolecules adsorption.
|
