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Titolo:
Probing DNA surface attachment and local environment using single moleculespectroscopy
Autore:
Osborne, MA; Barnes, CL; Balasubramanian, S; Klenerman, D;
Indirizzi:
Univ Cambridge, Dept Chem, Cambridge CB2 1EW, England Univ Cambridge Cambridge England CB2 1EW hem, Cambridge CB2 1EW, England
Titolo Testata:
JOURNAL OF PHYSICAL CHEMISTRY B
fascicolo: 15, volume: 105, anno: 2001,
pagine: 3120 - 3126
SICI:
1520-6106(20010419)105:15<3120:PDSAAL>2.0.ZU;2-L
Fonte:
ISI
Lingua:
ENG
Soggetto:
GREEN FLUORESCENT PROTEIN; RESONANCE ENERGY-TRANSFER; ROOM-TEMPERATURE; AQUEOUS-SOLUTION; CONFORMATIONAL TRANSITIONS; DYNAMICS; MICROSCOPY; FLUOROPHORES; EXCITATION; DIFFUSION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Physical, Chemical & Earth Sciences
Citazioni:
41
Recensione:
Indirizzi per estratti:
Indirizzo: Balasubramanian, S Univ Cambridge, Dept Chem, Lensfield Rd, Cambridge CB2 1EW, England Univ Cambridge Lensfield Rd Cambridge England CB2 1EW
Citazione:
M.A. Osborne et al., "Probing DNA surface attachment and local environment using single moleculespectroscopy", J PHYS CH B, 105(15), 2001, pp. 3120-3126

Abstract

We have employed single molecule fluorescence spectroscopy, using a total internal reflection geometry and wide-angle detection, to study the attachment of singly fluorescently labeled DNA to a silica surface by either a streptavidin-biotin of a covalent linkage. In both cases the DNA is highly monodispersed with no evidence for aggregation. The covalent coupling gave higher signal-to-noise than the streptavidin-biotin Linkage and was therefore studied in more detail. Two components in the photobleaching times, corresponding to different states of the tetramethyl rhodamine probe, were observed: a short and long component with populations in the ratio 6.7:1. Only rarely was interconversion between these two states detected during the 30-s observation time of the experiment. Hybridization experiments using a complementary strand of DNA labeled with a different fluorophore gave a low levelof colocalized fluorescence, indicating a significant fraction of the surface attached DNA was not available for hybridization. These results are consistent with the surface attached DNA spending significant time collapsed on the surface.

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Documento generato il 02/04/20 alle ore 06:21:56