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Team:Glendale CC AZ/Protocols/Ligation
From 2013.igem.org
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| - | + | ==== Ligation Protocol ==== | |
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| - | === Ligation Protocol === | + | |
Protocol from: http://parts.igem.org/Help:Protocols/Ligation | Protocol from: http://parts.igem.org/Help:Protocols/Ligation | ||
| - | === Materials === | + | ==== Materials ==== |
-digested plasmid backbone | -digested plasmid backbone | ||
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-thermal cycler | -thermal cycler | ||
| - | === Procedure === | + | ==== Procedure ==== |
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1. Add 2µL of digested plasmid backbone | 1. Add 2µL of digested plasmid backbone | ||
Revision as of 02:07, 23 September 2013
Glendale Community College Arizona
Protocols
Growth Curve Assay
NaCl Growth Curve Assay
Survival Growth Assay
Alkaline Lysis Plasmid Miniprep
Restriction Digest
DNA Isolation
Bioinformatics
Ligation
Transformation
Ligation Protocol
Protocol from: http://parts.igem.org/Help:Protocols/Ligation
Materials
-digested plasmid backbone -equimolar amount digested Part 1 -equimolar amount digested Part 2 -T4 DNA ligase buffer -T4 DNA ligase -dH2O -thermal cycler
Procedure
1. Add 2µL of digested plasmid backbone
2. Add equimolar amount of digested Part 1
3. Add equimolar amount of digested Part 2
4. Add 1µL T4 DNA ligase buffer.
5. Add 0.5µL T4 DNA ligase.
6. Add water to 10µL.
7. Place in pre-programmed thermal cycler at 16°C for 30 minutes then at 80°C for 20 minutes.
