Team:Glendale CC AZ/Protocols/Ligation
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+ | <h4>Glendale Community College Arizona<img | ||
+ | style="width: 200px; height: 58px;" alt="GCC" | ||
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+ | |||
+ | |||
+ | <h2><p>Protocols</p></h2><img style="border: 0px solid ; width: 400px; height: 200px;" alt="iGEM" src="https://static.igem.org/mediawiki/igem.org/1/19/Tubes2GCC.JPG" align="right"> | ||
+ | |||
+ | <a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/GrowthCurve">Growth Curve Assay</a></p> | ||
+ | <a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/NaCl">NaCl Growth Curve Assay</a></p> | ||
+ | <a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/SurivalGrowth">Survival Growth Assay</a></p> | ||
+ | <a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/AlkalineLysis">Alkaline Lysis Plasmid Miniprep </a></p> | ||
+ | <a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/RestrictionDigest">Restriction Digest</a></p> | ||
+ | <a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/DNAIsolation">DNA Isolation</a></p> | ||
+ | <a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/Bioinformatics">Bioinformatics</a></p> | ||
+ | <a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/Ligation">Ligation</a></p> | ||
+ | <a href="https://2013.igem.org/Team:Glendale_CC_AZ/Protocols/Transformation">Transformation</a></p> | ||
+ | |||
+ | |||
+ | <!--/.content-area--> | ||
+ | </body> | ||
+ | </html> | ||
+ | |||
+ | ==== 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. |
Latest revision as of 02:36, 28 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 TransformationLigation 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.