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Team:Aberdeen Scotland/Cloning/
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==Roche: Rapid DNA Ligation Kit== | ==Roche: Rapid DNA Ligation Kit== | ||
| - | Preparations: Prepare 1x conc DNA dilution buffer. | + | Preparations: Prepare 1x conc. DNA dilution buffer. |
1. Dissolve Vector DNA and Insert DNA, in carefully mixed DNA dilution buffer. <br> | 1. Dissolve Vector DNA and Insert DNA, in carefully mixed DNA dilution buffer. <br> | ||
Revision as of 19:46, 18 September 2009
University of Aberdeen - Pico Plumber
iGEM 2009
Notes about Cloning DNA
Molecular Cloning of DNA involves a number of steps, which can be broadly categorized as Restriction Digestion, Ligation and Transformation. Since DNA Digestion and Transformation are covered separately this page simply outlines the protocol used for Ligation.
Roche: Rapid DNA Ligation Kit
Preparations: Prepare 1x conc. DNA dilution buffer.
1. Dissolve Vector DNA and Insert DNA, in carefully mixed DNA dilution buffer.
2. Carefully mix T4 DNA Ligation buffer and subsequently add 10ul of DNA ligation buffer to mixture.
3. Add 1ul of T4 DNA Ligase to the mixture.
4. Incubate mixture for 5 minutes at 15-25*C
6. Store Ligation Mixture WITHOUT Heat Inactivating the Enzyme at -15 to -25 *C
 Protocols
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Competency 
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