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BP: Fachverband Biologische Physik

BP 19: Poster VII

BP 19.18: Poster

Dienstag, 17. März 2020, 14:00–16:00, P2/3OG

High-yield fabrication of DNA and RNA constructs for single molecule force and torque spectroscopy experiments. — •Flavia Stal Papini, Mona Seifert, and David Dulin — Junior Research Group 2, Interdisciplinary Center for Clinical Research, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Cauerstraße 4, 91058 Erlangen, Germany

Single molecule biophysics experiments have enabled the observation of biomolecules with a great deal of precision in space and time, e.g. nucleic acids mechanical properties and protein*nucleic acid interactions using force and torque spectroscopy techniques. The success of these experiments strongly depends on the efficient design and fabrication of complex nucleic acid structures, as the outcome of the experiment strongly depends on the high quality of the final construct. Though the molecular biology techniques involved are well known, the fabrication of nucleic acid constructs for single molecule experiments still remains a difficult task. We developed new protocols to generate high-yield and high-quality coilable double-stranded DNA and RNA constructs, as well as DNA and RNA hairpins with *500*1000 bp long stems (Papini et al., NAR 2019). A new approach based on single-stranded DNA annealing is presented and its efficiency in the fabrication of complex DNA constructs is shown in magnetic tweezers assays. Our protocols enable the design of a large range of nucleic acid constructs for single molecule biophysics experiments.