Millisecond analysis of double stranded DNA with fluorescent intercalator by micro-thermocontrol-device

TALANTA

卷:79

Article

Arata, Hideyuki F.^1,2  Gillot, Frederic^1,3  Collard, Dominique^1,3  Fujita, Hiroyuki¹ 

[1] Univ Tokyo, IIS, Meguro Ku, Tokyo 1538505, Japan

[2] Japan Soc Promot Sci, Tokyo, Japan

[3] CNRS, LIMMS, IIS, F-75700 Paris, France

关键词: Intercalator;    DNA;    Denaturation;    Microheater;    Millisecond;   

DOI  :  10.1016/j.talanta.2009.04.045

来源: Elsevier

【 摘 要 】

Study of interaction between DNA and intercalator at molecular level is important to understand the mechanisms of DNA replication and repair. A micro-fabricated local heating thermodevice was adapted to perform denaturation experiments of DNA with fluorescent intercalator on millisecond time scale. Response time of complete unzipping of double stranded DNA, 16 mu m in length, was measured to be around 5 min by commercial thermocycler. Response time of quenching of double stranded DNA with fluorescent intercalator SYBR Green was measured to be 10 ms. Thus, quenching properties owing to strand unzipping and denaturation at base pair level were distinguished. This method has provided easy access to measure this parameter and may be a powerful methodology in analyzing biomolecules on millisecond time scale. (C) 2009 Elsevier B.V. All rights reserved.

【 授权许可】

Free