TY  - JOUR
A1  - Linck, Lena
A1  - Reiss, Edda
A1  - Bier, Frank Fabian
A1  - Resch-Genger, Ute
T1  - Direct labeling rolling circle amplification as a straightforward signal amplification technique for biodetection formats
JF  - Analytical methods : advancing methods and applications
N2  - Biodetection formats, such as DNA and antibody microarrays, are valuable tools in the life sciences, but for some applications, the detection limits are insufficient. A straightforward strategy to obtain signal amplification is the rolling circle amplification (RCA), an easy, isothermal, and enzymatic nucleic acid synthesis that has already been employed successfully to increase the signal yield for several single-analyte and multiplexing assays in conjunction with hybridization probes. Here, we systematically investigated the parameters responsible for the RCA driven signal amplification with fluorescent labels, such as the type of fluorophore chosen, labeling strategy, composition of reaction solution, and number of handling steps. In labeling strategies, post-synthetic labeling via a Cy3-hybridization probe was compared to the direct incorporation of fluorescent Cy3-dUTP and DY-555-dUTP into the nascent strand during synthesis. With our direct labeling protocol, the assay's runtime and handling steps could be reduced while the signal yield was increased. These features are very attractive for many detection formats but especially for point-of-care diagnostic kits that need to be simple enough to be performed by scientifically untrained personnel.
Y1  - 2012
U6  - https://doi.org/10.1039/c2ay05760c
SN  - 1759-9660
VL  - 4
IS  - 5
SP  - 1215
EP  - 1220
PB  - Royal Society of Chemistry
CY  - Cambridge
ER  - 
TY  - JOUR
A1  - Reiss, Edda
A1  - Hoelzel, Ralph
A1  - Bier, Frank Fabian
T1  - Synthesis and stretching of rolling circle amplification products in a flow-through system
N2  - Enzymatic isothermal rolling circle amplification (RCA) produces long concatemeric single-stranded DNA (ssDNA) molecules if a small circular ssDNA molecule is applied as the template. A method is presented here in which the RCA reaction is carried out in a flow-through system, starting from isolated surface-tethered DNA primers. This approach combines gentle fluidic handling of the single-stranded RCA products, such as staining or stretching via a receding meniscus, with the option of simultaneous (fluorescence) microscopic observation. It is shown that the stretched and surface-attached RCA products are accessible for hybridization of complementary oligonucleotides, which demonstrates their addressability by complementary base pairing. The long RCA products should be well suited to bridge the gap between biomolecular nanoscale building-blocks and structures at the micro- and macroscale, especially at the single- molecule level presented here.
Y1  - 2009
UR  - http://www3.interscience.wiley.com/cgi-bin/jhome/107640323
U6  - https://doi.org/10.1002/smll.200900319
SN  - 1613-6810
ER  - 
TY  - THES
A1  - Reiß, Edda
T1  - Erzeugung einzelsträngiger DNA-Gerüststränge mittels Rolling-Circle-Amplifikation für den Einsatz in der Nanobiotechnologie
Y1  - 2009
CY  - Potsdam
ER  -