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TPMProtocol - Bi1X Single molecule techniques The tethered...

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Bi1X: Single molecule techniques The tethered particle method In this lab we will visualize single DNA molecules bound to a 0.5µm bead. This DNA will have a binding site for the restriction enzyme KpnI. When flowing in this enzyme we’ll see its effects by counting the tethers that get lost due to the DNA getting cut. You will be provided with flow chambers with the single DNA molecules already bound to the surface and to the beads. Your job is to mount the sample on the microscope, find the beads and take a movie of the beads while flowing in the restriction enzyme. The flow chambers: A lot of things happened behind the scenes. First, let’s familiarize ourselves with the flow chambers. The chamber is made out of a microscope slide that has two holes drilled. Double sticky tape is sandwiched between this slide and a microscope coverslip. The tape is cut such that a little chamber is created between the coverslip and the slide. Tubing is connected to the wholes using Epoxy.
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