Copper-free click chemistry for attachment of biomolecules in magnetic tweezers

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• 作者：Jorine M. Eeftens ; Jaco van der Torre ; Daniel R. Burnham ; Cees Dekker
• 关键词：Magnetic tweezers ; Copper ; free click chemistry ; SPAAC reactions ; Surface chemistry ; DNA immobilization
• 刊名：BMC Biophysics
• 出版年：2015
• 出版时间：December 2015
• 年：2015
• 卷：8
• 期：1
• 全文大小：1,317 KB
• 刊物主题：Biophysics and Biological Physics;
• 出版者：BioMed Central
• ISSN：2046-1682

文摘

Background Single-molecule techniques have proven to be an excellent approach for quantitatively studying DNA-protein interactions at the single-molecule level. In magnetic tweezers, a force is applied to a biopolymer that is anchored between a glass surface and a magnetic bead. Whereas the relevant force regime for many biological processes is above 20pN, problems arise at these higher forces, since the molecule of interest can detach from the attachment points at the surface or the bead. Whereas many recipes for attachment of biopolymers have been developed, most methods do not suffice, as the molecules break at high force, or the attachment chemistry leads to nonspecific cross reactions with proteins.