Millisecond analysis of double stranded DNA with fluorescent intercalator by micro-thermocontrol-device.
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 microm 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.
Arata
Hideyuki F
HF
Institute of Industrial Science (IIS), The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8505, Japan. arata@sanken.osaka-u.ac.jp
Gillot
Frederic
F
Collard
Dominique
D
Fujita
Hiroyuki
H
D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000378:metabolism*; D007364:Intercalating Agents / Q000378:metabolism*; D009691:Nucleic Acid Denaturation; D013696:Temperature; D013997:Time Factors
D005456:Fluorescent Dyes; D007364:Intercalating Agents; D004247:DNA