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Team:SDU-Denmark/Tools
From 2009.igem.org
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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? | ||
| + | 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.
