Koch Lab:Protocols/Optical Tweezers: Difference between revisions
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(New page: ==Background== ==Setup== ===Basic Setup=== ===Our Setup=== ===Future Improvements=== ==Calibration== ==Experiments==) |
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==Calibration== | ==Calibration== | ||
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There are numerous experiments that could be performed using an Optical Tweezers. KochLab uses it for biological applications. And we separate the experiments into two regimes: '''DNA Applications''' and '''Kinesin Applications'''. | |||
===DNA Experiments=== | |||
#DNA Stretching - The essence of this technique is to stretch DNA. It is possible to label DNA so with biotin (for adhesion to a streptavidin coated microsphere) and digoxigenin (for adhesion to anti-digoxigenin). These labels allow for the creation of dsDNA tethers because of the nature of anti-dig bonding with glass. The microsphere allows our OT to probe the sample and stretch the DNA. From here there are many avenues to take. First you can reproduce worm-like chain analysis. You could also perform some protein-DNA analysis and observe the amount of force required to remove proteins bound to dsDNA. There are also experiments investigating the nature of DNA intercalaters enhanced by stretching. | |||
#DNA Unzipping - Steve created an unzipping construct | |||
#Transcription - | |||
===Kinesin Experiments=== | |||
#Bead Motility Assay - | |||
Revision as of 10:06, 23 October 2009
Background
Setup
Basic Setup
Our Setup
Future Improvements
Calibration
Experiments
There are numerous experiments that could be performed using an Optical Tweezers. KochLab uses it for biological applications. And we separate the experiments into two regimes: DNA Applications and Kinesin Applications.
DNA Experiments
- DNA Stretching - The essence of this technique is to stretch DNA. It is possible to label DNA so with biotin (for adhesion to a streptavidin coated microsphere) and digoxigenin (for adhesion to anti-digoxigenin). These labels allow for the creation of dsDNA tethers because of the nature of anti-dig bonding with glass. The microsphere allows our OT to probe the sample and stretch the DNA. From here there are many avenues to take. First you can reproduce worm-like chain analysis. You could also perform some protein-DNA analysis and observe the amount of force required to remove proteins bound to dsDNA. There are also experiments investigating the nature of DNA intercalaters enhanced by stretching.
- DNA Unzipping - Steve created an unzipping construct
- Transcription -
Kinesin Experiments
- Bead Motility Assay -
