Analysis of DNA coverage using enzymatic cleavage of fluorescent labels

Title
Analysis of DNA coverage using enzymatic cleavage of fluorescent labels
Authors
김미정정순김태송김상경
Keywords
DNA quantification; SAM; DNase 1; surface density; fluorescence; Gold; Biosensor; Deoxyribonuclease I; Fluorescent label
Issue Date
2011-01
Publisher
BioChip Journal
Citation
VOL 5, NO 1, 39-46
Abstract
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.
URI
http://pubs.kist.re.kr/handle/201004/40065
ISSN
1976-0280
Appears in Collections:
KIST Publication > Article
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