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Team:SDU-Denmark/Tools
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=Plasmid ligation helper= | =Plasmid ligation helper= | ||
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| - | To avoid having to manually calculate how | + | To avoid having to manually calculate how many uL's of DNA and plasmid we should use to achieve optimal ligation circumstances, we created a small Program for the texas instruments calculators. It's simple called Ligation, and you can get it here. |
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| - | Q7: How much DNA should be used in a ligation using T4 DNA Ligase? | + | <font size=1px>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. | + | 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. </font> From [http://www.neb.com/nebecomm/products/faqproductM0202.asp T4 DNA Ligase FAQ] |
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Revision as of 06:22, 21 October 2009
Plasmid ligation helper
To avoid having to manually calculate how many uL's of DNA and plasmid we should use to achieve optimal ligation circumstances, we created a small Program for the texas instruments calculators. It's simple called Ligation, and you can get it here.
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. From T4 DNA Ligase FAQ
