During the early stages of this Workpackage a series of key decisions will have been made in other Workpackages that will influence the nature of the work to be carried out.  (i) A decision will have been made as to which of the various molecular motors will provide the best means for moving a magnetic bead.  (ii) A decision about the size and composition/nature of the most reliable and easily detected magnetic bead.  (iii) Development of a suitable surface on which the motor will function, with grooves or channels into which the motor and DNA can be localised.

Outcomes:

The ENS group has developed a micro-fluidic device made of PDMS (an elastomer), which defines the reaction chamber (channel) and is placed on a microscope coverslip, to allow for visualization under a microscope. Appropriate tubing on the PDMS allow for easy exchange of buffers. The reaction cell thus defined is easy and cheap and can be made in many exemplars.

We have investigated mechanism for motor immobilisation and release as part of the Proof of Principle for the Mol Switch Device Patent Application.

We have coupled the manipulation of DNA using the magnetic trap set-up with Total Internal Reflection Microscopy (TIRM) that allows for the observation by fluorescence of  protein interacting with DNA. We are using that set-up to better study and characterize  our motors. We are also planning of using TIRM as a method to detect the motion of the bead which could be directly read by a photosensor (thus avoiding the requirement of an optical microscope system) and allowing for a device that will couple directly the enzymatically induced motion of the bead to an electric signal.