Monitoring the amplification of CATCH, a 3SR based cooperatively coupled isothermal amplification system, by fluorimetric methods.
Three different types of fluorescence detection methods were employed to monitor amplification of a previously established isothermal cooperatively coupled amplification system as it can serve as a tool for the investigation of fundamental issues in evolutionary optimization. By using 5'IRD-41 fluorescent labeled primers, the intercalating dye TOPRO-1 and a 5'fluorescin/3'DABCYL 4-(4-dimethylamino-phenylazo)benzoic acid labeled ss 24 nt DNA, evolving molecular cooperation is accessible, sequence specifically as well as non-sequence-specifically without using radioactivity.
Ehricht
R
R
Department of Molecular Information Processing, Institute of Molecular Biotechnology, Beutenbergstrasse 11, 07745 Jena, Germany. rehr@imb-jena.de
Kirner
T
T
Ellinger
T
T
Foerster
P
P
McCaskill
J S
JS
D004247:DNA / Q000032:analysis; D017931:DNA Primers; D005456:Fluorescent Dyes; D005470:Fluorometry / Q000379:methods*; D005784:Gene Amplification; D007364:Intercalating Agents; D009690:Nucleic Acid Conformation; D017422:Sequence Analysis, DNA; D004124:p-Dimethylaminoazobenzene / Q000031:analogs & derivatives
D017931:DNA Primers; D005456:Fluorescent Dyes; D007364:Intercalating Agents; C063127:4-(4-dimethylaminophenylazo)benzoic acid; D004247:DNA; D004124:p-Dimethylaminoazobenzene