Medizinische Universität Graz - Research portal
Selected Publication:
FoldesPapp, Z; Thyberg, P; Bjorling, S; Holmgren, A; Rigler, R.
Exonuclease degradation of DNA studied by fluorescence correlation spectroscopy
NUCLEOS NUCLEOT. 1997; 16(5-6): 781-787.
Doi: 10.1080/07328319708002951
Web of Science
FullText
FullText_MUG
- Leading authors Med Uni Graz
- Földes-Papp Zeno
- Altmetrics:
- Dimensions Citations:
- Plum Analytics:
- Scite (citation analytics):
- Abstract:
- Here we developed an accurate method for kinetic analysis of enzymatic degradation processes of double and/or single-stranded DNA/oligonucleotides using fluorescent reporter dyes. 217-bp DNA fragments were produced by polymerase chain reaction and cleaved by the 3' to 5' exonuclease activity of T7-DNA polymerase. The analysis of the products was performed by Fluorescence Correlation Spectroscopy measuring autocorrelation amplitudes and diffusion times. We give proof of: (i) complete enzymatic degradation, (ii) retardation of complete enzymatic degradation by internally labelled Rhodamine-4-nucleotides and Cy5-nucleotides, respectively. Data evaluation by global analysis indicated first-order reaction kinetics with full-length DNA and free fluorescent nucleotides in the time window of measurements used.