Evaluating Your Thermal Denaturation Data - Agilent Technologies 8453 Operator's Manual

• Enter the DNA length of the sample in base pairs (bp) in the DNA Length
• You can enter a comment (for example a description of the sample) in the
Press the Run button in the Sample start the acquisition and to display the All
Spectra window, the Trace window and the Sample Information window. First
the Peltier temperature controller sets the starting temperature at the cell
holder. After the hold time which has been defined in the Temperature Ramp
dialog box the first spectrum is measured and the Peltier temperature
controller sets the next temperature of the ramp.
During the thermal measurement you can follow the acquisition of the spectra
and the temperature traces in real-time in the All Spectra window and in the
Trace window.
After the thermal denaturation measurement has been finished or aborted by
the user, the data are stored as specified in the Temperature & Options dialog
box. The traces are evaluated by the procedure, that has been defined in the
Calculation Parameters dialog box. These results are displayed in the Result
window.

Evaluating your Thermal Denaturation Data

After the thermal denaturation run has been finished you have the possibility
to reevaluate the experimental data. Since full spectra have been acquired
during the measurement, all parameters in the Calculation Parameters dialog
box can be changed. Click OK to make the changes active. The new results are
displayed in the Results of Heating Trace window. If the method parameter
has been changed the modified flag Mod will appear in the tool bar of the
graphical user interface.
When you press the Edit Sample Information button in the from the Results of
Heating Trace window, the Sample Information dialog box is displayed. It
enables you to include information on your samples and for use in results
calculations.
Biochemical Analysis Software for Agilent ChemStation
(K) field. The DNA length is included in the Results of Trace table and
reports and is used in the %G-C calculation.
Comment field.
Thermal Denaturation Mode 3
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