Melting Temperature Tm of Nucleic Acids

LifePower TM-Light is a software program designed to acquire and store data for the evaluation of the meltpoint of a nucleic acid sample. Data generated with this software package can be saved, printed or exported in ASCII or EXCEL format to be used for further calculations.

Abs vs. temperature data with autoscaled graphs and real-time data storage are provided.
User defined comments are stored within the data file and all entered parameters are saved as customized command file or as default parameters for the next experiment.
Once the thermal experiment starts, LifePower TM-light continuously acquires the Abs and the Temperatures at regular user preset time or temperature interval until the final temperature has been reached. Regular time interval should be preferred because it allows further kinetic analysis if possible.
Real time graphics are automatically displayed and updated during data acquisition. At the end of the experiment, the displayed raw data (by default) that can be proceeded and normalized.
The melting temperature (Tm) value and the shape of the thermal transition depends on the sequence and length of the nucleic acids as well as experimental conditions (ionic strength, pH, etc). The Tm value is a transition point where the complex is half-dissociated, or half-formed. This transition point corresponds approximately to the inflection point of the melting curve where the slope becomes steepest.

DNA thermal melting curve will provide with not only the melting temperature (Tm), but also the free energy and its associated enthalpy and entropy contributions.

2D-Data Acquisition


The basic feature in the nucleic acid thermal analysis is the 2D Data Acquisition, where the experiment is executed in fixed wavelength/wavelength program mode (generally between 220 nm and 330 nm and up to 6 selectable wavelengths) at regular user preset time or temperature interval until the final temperature has been reached. The raw or baseline-corrected data are displayed in 2D mode (Abs, temperature with different color curves for each wavelength in one diagram).

3D-Data Acquisition


A new feature in the nucleic acid thermal analysis is the 3D SCAN, where the experiment is executed in spectrum scan mode (generally between 220 nm and 330 nm) at regular user preset time or temperature interval until the final temperature has been reached. The raw or baseline-corrected data are displayed in 3D mode (Abs, wavelength, temperature). It is also possible to obtain a 2D (Abs, temperature) thermal transition curve at any wavelength.
3D Scan is particularly useful at the beginning of a new research project, since it helps to detect whether the melting process involves only two or further more components. For the complicated reactions where there are more than one equilibrium, it is possible to identify the component by spectral deconvolution, and separate different thermal transitions by setting the appropriate wavelength.