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Example for Hyb-probe improvement using MeltCalc©The ultimate thermodynamic modelling spreadsheet for ExcelThe following
LightCycler melting curves have been achieved before and after thermodynamic
Hyb-Probe assay optimization using MeltCalc© Professional which runs
as Excel© "Add-In" in almost every Windows environment. Excel
7.0 or higher is required for execution. For this example
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First probe set, manually selected | Second probe set, selected with MeltCalc | |||||||||||||||||||||||||||||||||||||||
Prediction
of melting temperatures (Tm) with MeltCalc© The ultimate thermodynamic modelling spreadsheet for Excel |
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This correlation was calculated from 35 independent assays from us (n=25) and from data in the literature (n=10). All melting temperature were measured in LightCycler hybridization probe assays. | ||||||||||||||||||||||||||||||||||||||||
New
features of this version are: Calculation of Tm now including "dangling end"parameters (if applicable). Maximum possible sequence length increased from 256bp to 1755bp for all features. Maximum possible alignment for cross hybridization is shown graphically. Mutations can be inserted by bp and base exchange. A formula for calculation of Tm for long sequences is added . Probe design: Minor modifications: |
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First
page: Cross-Hybridization Tms are highlighted using color coding in dependence of their hybridization temperature |
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Second
page: Cross-Hyb are shown graphically (partial display) |
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Mutations can be inserted by bp and base exchange | ||||||||||||||||||||||||||||||||||||||||