Highly parallel magnetic tweezers by targeted DNA tethering.
Single-molecule force-spectroscopy methods such as magnetic and optical tweezers have emerged as powerful tools for the detailed study of biomechanical aspects of DNA-enzyme interactions. As typically only a single molecule of DNA is addressed in an individual experiment, these methods suffer from a low data throughput. Here, we report a novel method for targeted, nonrandom immobilization of DNA-tethered magnetic beads in regular arrays through microcontact printing of DNA end-binding labels. We show that the increase in density due to the arrangement of DNA-bead tethers in regular arrays can give rise to a one-order-of-magnitude improvement in data-throughput in magnetic tweezers experiments. We demonstrate the applicability of this technique in tweezers experiments where up to 450 beads are simultaneously tracked in parallel, yielding statistical data on the mechanics of DNA for 357 molecules from a single experimental run. Our technique paves the way for kilo-molecule force spectroscopy experiments, enabling the study of rare events in DNA-protein interactions and the acquisition of large statistical data sets from individual experimental runs.
Kavli Institute of Nanoscience, Delft University of Technology, Delft, Lorentzweg 1, 2628 CJ, The Netherlands.
Iwijn
I
De Vlaminck
Thomas
T
Henighan
Marijn T J
MT
van Loenhout
Indriati
I
Pfeiffer
Julius
J
Huijts
Jacob W J
JW
Kerssemakers
Allard J
AJ
Katan
Anja
A
van Langen-Suurling
Emile
E
van der Drift
Claire
C
Wyman
Cees
C
Dekker
D001696:Biomechanical Phenomena; D004247:DNA / Q000737:chemistry*; D059346:Magnets / Q000737:chemistry*; D018625:Microscopy, Atomic Force / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis; D052898:Optical Tweezers
D004247:DNA