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
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.