Difference between revisions of "DNA Melting: Model function and parameter estimation by nonlinear regression"
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===Assumptions=== | ===Assumptions=== | ||
* The dye is divided into two populations, bleached and unbleached, with concentrations S and <span style="text-decoration: overline">S</span> | * The dye is divided into two populations, bleached and unbleached, with concentrations S and <span style="text-decoration: overline">S</span> | ||
| + | ** The initial dye concentration is S + <span style="text-decoration: overline">S</span> | ||
* Bleached and unbleached molecules bind to dsDNA with the same affinity. | * Bleached and unbleached molecules bind to dsDNA with the same affinity. | ||
* Only bound dye molecules may be excited. | * Only bound dye molecules may be excited. | ||
Revision as of 17:32, 13 November 2012
This subject matter will be explained in lecture, in tutorials, and in the lab.
Photobleaching model
Assumptions
- The dye is divided into two populations, bleached and unbleached, with concentrations S and S
- The initial dye concentration is S + S
- Bleached and unbleached molecules bind to dsDNA with the same affinity.
- Only bound dye molecules may be excited.
- An excited fluorophore will either bleach with probability p or return to the ground state with probability 1-p.
- Mechanisms for returning to the ground state include fluorescence, phosphorescence, and non-radiative relaxation.
- Only excited fluorophore molecules bleach
- Bleaching is irreversible
