DNA Transcription

Scientists can use optical tweezers to trap beads, as depicted at the right, and catch a biomolecule, such as DNA, in between. This biomolecule can then be manipulated by moving the beads, while the force and extension are measured. Fluorescently labeled proteins can be visualized with confocal or STED fluorescence microscopy. The combination of optical tweezers with simultaneous multicolor fluorescence measurements allows correlating the mechanical properties of the DNA with the protein activity. With optical tweezers – fluorescence microscopy you can:

• Visualize and directly measure DNA-protein interactions in DNA transcription to find the kinetics and exact mechanisms involved at the single-molecule level
• Correlate the mechanical properties of the DNA with the binding location and quantity of DNA replication proteins
• Probe and monitor the activity and states of motor proteins, such as RNA polymerase, and extract novel information on the single stepping of biomolecular motors and their enzymatic mechanisms with basepair resolution
• Investigate the conformational states of DNA or RNA molecules and correlate them with transcription control
• Perform experiments under biologically relevant conditions and highly crowded environments and link the in vitro experiment with the in vivo situation
• Study the effect of small molecules and biologics in DNA transcription pathways.

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Optical Tweezers and Fluorescence Microscopy

Scientists can use acoustic forces to manipulate, stretch and measure hundreds of single molecules at the same time. Multiple DNA, RNA or protein-coated molecules can be tethered between a bead and a glass surface. Using Acoustic Force Spectroscopy (AFS) technology you can then apply controlled forces on all beads synchronously and probe the mechanical properties of each molecule in parallel. With this highly parallel single-molecule method you can:

• Investigate the mechanics of many DNA molecules in the presence of DNA transcription proteins and in different buffers, at the single-molecule level
• Gain insights into the dynamics and cooperative behavior of protein binding to the DNA
• Measure the enzymatic activity, kinetics, and stepping mechanisms of hundreds of individual motor proteins in parallel
• Perform experiments under biologically relevant conditions and link the in vitro experiment with the in vivo
• Study the effect of small molecules and biologics in DNA transcription pathways.
• Receive large datasets containing many single-molecule measurements from a single experiment.

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Parallel Single-Molecule Force Spectroscopy