Print Email Facebook Twitter Characterizing single-molecule dynamics of viral RNA-dependent RNA polymerases with multiplexed magnetic tweezers Title Characterizing single-molecule dynamics of viral RNA-dependent RNA polymerases with multiplexed magnetic tweezers Author Kuijpers, L.C. (TU Delft BN/Nynke Dekker Lab) van Laar, T. (TU Delft BN/Nynke Dekker Lab) Janissen, R. (TU Delft BN/Cees Dekker Lab) Dekker, N.H. (TU Delft BN/Nynke Dekker Lab) Date 2022 Abstract Multiplexed single-molecule magnetic tweezers (MT) have recently been employed to probe the RNA synthesis dynamics of RNA-dependent RNA polymerases (RdRp). Here, we present a protocol for simultaneously probing the RNA synthesis dynamics of hundreds of single polymerases with MT. We describe the preparation of a dsRNA construct for probing single RdRp kinetics. We then detail the measurement of RdRp RNA synthesis kinetics using MT. The protocol is suitable for high-throughput probing of RdRp-targeting antiviral compounds for mechanistic function and efficacy. For complete details on the use and execution of this protocol, please refer to Janissen et al. (2021). Subject BiophysicsMicrobiologyMolecular biologyMolecular/Chemical probesSingle-molecule assays To reference this document use: http://resolver.tudelft.nl/uuid:40a6da94-63e6-495a-a9f1-205d995331cf DOI https://doi.org/10.1016/j.xpro.2022.101606 Source STAR Protocols, 3 (3) Part of collection Institutional Repository Document type journal article Rights © 2022 L.C. Kuijpers, T. van Laar, R. Janissen, N.H. Dekker Files PDF 1_s2.0_S2666166722004865_main.pdf 4.26 MB Close viewer /islandora/object/uuid:40a6da94-63e6-495a-a9f1-205d995331cf/datastream/OBJ/view