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Volumn 4, Issue 4, 2008, Pages 744-747

Poly(l-lysine)-graft-dextran copolymer accelerates DNA hybridization by two orders

Author keywords

[No Author keywords available]

Indexed keywords

COPOLYMERS; IONIC CONDUCTION;

EID: 41149089304     PISSN: 1744683X     EISSN: 17446848     Source Type: Journal    
DOI: 10.1039/b717478k     Document Type: Article
Times cited : (33)

References (33)
  • 23
    • 84943009565 scopus 로고    scopus 로고
    • note
    • A TAMRA-labeled DNA (T25P:1 pmol) was incubated at 20 °C in PBS buffer (10 mM sodium phosphate, 0.5 mM EDTA, 150 mM NaCl, pH 7.2) in the absence or presence of cationic copolymer (copolymer-DNA charge ratio = 10). Then, its complementary DABCYL-labeled DNA (D25T:1 pmol) was added to initiate DNA hybridization. The change in fluorescence intensity of the mixture (total volume 2 ml) in a 10 mm2 glass cuvette was recorded on a Jasco FP-6500 spectrofluorometer equipped with a peltier thermostatically controlled cell holder at excitation and emission wavelength of 540 and 570 nm, respectively, with excitation and emission slits at 10 nm. Each experiment was repeated at least twice. The value of fluorescence intensity was calculated with the following equation to yield a rate constant of the second-order reaction. kt = (I0 - It)/[CDNA (It - I )]. Where I0 is the initial fluorescence intensity of the TAMRA-labeled DNA, It is that at time t, and I is that after the reaction reached equilibrium. The value of Iwas practically obtained by measuring the mixture that had been heat-treated (heating at 90 °C for 5 min, followed by slow cooling to 20 °C). CDNA is the initial concentration of DNA strands. The procedure was the same in the reaction of a couple of complementary 20 mer DNAs, T20P and D20T, but at final conc. = 2 nM


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