Team:SDU-Denmark/Tools

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=Plasmid ligation helper=
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To avoid having to manually calculate how much DNA and how much plasmid we should use to achieve such the above requirements, we created a small Program for the texas instruments calculators.
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Q7: How much DNA should be used in a ligation using T4 DNA Ligase?
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A7: The unit definition uses 0.12 μM (300 μg/ml) lambda HindIII fragments. The high DNA concentration can be used for linker ligation. To promote circle formation, useful in transformation, a lower total DNA concentration should be used. The overall concentration of vector + insert should be between 1-10 μg/ml for efficient ligation. Insert:vector molar ratios between 2 and 6 are optimal for single insertions. Ratios below 2:1 result in lower ligation efficiency. Ratios above 6:1 promote multiple inserts. If you are unsure of your DNA concentrations, perform multiple ligations with varying ratios.
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Revision as of 06:18, 21 October 2009






Plasmid ligation helper

To avoid having to manually calculate how much DNA and how much plasmid we should use to achieve such the above requirements, we created a small Program for the texas instruments calculators.



Q7: How much DNA should be used in a ligation using T4 DNA Ligase?

A7: The unit definition uses 0.12 μM (300 μg/ml) lambda HindIII fragments. The high DNA concentration can be used for linker ligation. To promote circle formation, useful in transformation, a lower total DNA concentration should be used. The overall concentration of vector + insert should be between 1-10 μg/ml for efficient ligation. Insert:vector molar ratios between 2 and 6 are optimal for single insertions. Ratios below 2:1 result in lower ligation efficiency. Ratios above 6:1 promote multiple inserts. If you are unsure of your DNA concentrations, perform multiple ligations with varying ratios.