| Info. | Vol.5 - No.1 (2011.03.20) |
|---|---|
| Title | Analysis of DNA coverage using enzymatic cleavage of fluorescent labels |
| Authors | Mi Jung Kim1,??/sup>, Shun Zheng1,??/sup>, Tae Song Kim1 & Sang Kyung Kim1 |
| Institutions | 1Nano-Bio Center, Korea Institute of Science and Technology (KIST), P.O. Box 131, Cheongryang, Seoul 130-650, Korea ??/sup>These authors contributed equally to this work Correspondence and requests for materials should be addressed to S.K. Kim( sangk@kist.re.kr) |
| Abstract | In this study, a method to determine the degree of DNA coverage on gold surface using a flu-orescent 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. |
| Keyword | DNA quantification, Gold, Biosensor, Deo-xyribonuclease I, Fluorescent label |
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