Analysis of DNA coverage using enzymatic cleavage of fluorescent labels
- Analysis of DNA coverage using enzymatic cleavage of fluorescent labels
- 김미정; 정순; 김태송; 김상경
- DNA quantification; SAM; DNase 1; surface density; fluorescence; Gold; Biosensor; Deoxyribonuclease I; Fluorescent label
- Issue Date
- BioChip Journal
- VOL 5, NO 1, 39-46
- In this study, a method to determine the
degree of DNA coverage on gold surface using a fluorescent
dye and Deoxyribonuclease I (DNase I) was
developed. Gold surfaces were first functionalized with
fluorescent dye labeled DNA, which was followed by
DNase I digestion. The number of released fluorescent
dyes represented the quantity of DNA immobilized.
The use of DNase I to enzymatically cleave the fluorescent
dye decreased the time needed for quantification
to 1 h. In this way, the effect of a back-filler, DNA concentration,
incubation time and spacer on DNA immobilization
via gold-thiol chemistry was examined.
The concentration of oligonucleotides and the ionic
environment were the major factors determining DNA
density. We also found that the length of DNA molecule
influenced the rate of immobilization on Au. In
addition, mixed self-assembled monolayer (mixed-
SAM) with various composition and the the effect on
DNA density was investigated.
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