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Team:Glendale CC AZ/Protocols/Ligation
From 2013.igem.org
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E.Soboslay (Talk | contribs) (→Procedure) |
E.Soboslay (Talk | contribs) (→Materials) |
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-digested plasmid backbone | -digested plasmid backbone | ||
| - | -equimolar amount | + | -equimolar amount digested Part 1 |
| - | -equimolar amount | + | -equimolar amount digested Part 2 |
-T4 DNA ligase buffer | -T4 DNA ligase buffer | ||
-T4 DNA ligase | -T4 DNA ligase | ||
-dH2O | -dH2O | ||
| + | -thermal cycler | ||
== Procedure == | == Procedure == | ||
Revision as of 22:58, 26 August 2013
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.
