Development of DNA methylation measurement device using nano-carbon film electrode
| Project/Area Number |
20350042
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Analytical chemistry
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| Research Institution | National Institute of Advanced Industrial Science and Technology |
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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)
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| 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)
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| Project Period (FY) |
2008 – 2010
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| Project Status |
Completed (Fiscal Year 2010)
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| Budget Amount *help |
¥19,630,000 (Direct Cost: ¥15,100,000、Indirect Cost: ¥4,530,000)
Fiscal Year 2010: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2009: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
Fiscal Year 2008: ¥8,190,000 (Direct Cost: ¥6,300,000、Indirect Cost: ¥1,890,000)
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| Keywords | メチル化DNA / チップ分析 / 電気化学検出 / センサ / 電気化学 / シトシン / ナノカーボン薄膜 / 酵素分解 / ECRスパッタ / パルスボルタンメトリ / DNA / メチル化 / オリゴヌクレオチド / 酵素 / 電気化学分析 / 表面置換基 |
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| 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.
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Report
(4 results)
Research Products
(50 results)
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[Presentation] DNAメチル化の電気化学検出2009
Author(s)
後藤圭佑, 加藤大, 関岡直行, 上田晃生, 栗田僚二, 廣野滋, 丹羽修
Organizer
日本分析化学会第58年会
Place of Presentation
北大
Year and Date
2009-09-25
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