Team:UGA-Georgia/Protocol
From 2013.igem.org
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1. Plasmid extraction: ZYMO RESEARCH | 1. Plasmid extraction: ZYMO RESEARCH | ||
[http://www.zymoresearch.com/media/downloads/47/D4036i.pdf] | [http://www.zymoresearch.com/media/downloads/47/D4036i.pdf] | ||
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2. PCR: KOD/ Phusion Polymerase (EMD Chemicals Inc.,NJ) (Novagen Protocol)/ NEB | 2. PCR: KOD/ Phusion Polymerase (EMD Chemicals Inc.,NJ) (Novagen Protocol)/ NEB | ||
Manufacturer’s kit | Manufacturer’s kit | ||
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3. Restriction Digestion: New England Biolabs (Beverly, MA) | 3. Restriction Digestion: New England Biolabs (Beverly, MA) | ||
Manufacturer’s kit | Manufacturer’s kit | ||
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4. Ligation: Rapid DNA Ligase (Roche Applied Science, Indianapolis, IN). | 4. Ligation: Rapid DNA Ligase (Roche Applied Science, Indianapolis, IN). | ||
Manufacturer’s kit | Manufacturer’s kit | ||
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5. Chemical competent cell and Transformation (Heat shock) | 5. Chemical competent cell and Transformation (Heat shock) | ||
(Protocol developed by hands on experience) | (Protocol developed by hands on experience) | ||
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6. PCR product purification kit and gel extraction kit (ZYMO RESEARCH) | 6. PCR product purification kit and gel extraction kit (ZYMO RESEARCH) |
Latest revision as of 18:24, 22 September 2013
Protocols
1. Plasmid extraction: ZYMO RESEARCH [http://www.zymoresearch.com/media/downloads/47/D4036i.pdf]
2. PCR: KOD/ Phusion Polymerase (EMD Chemicals Inc.,NJ) (Novagen Protocol)/ NEB Manufacturer’s kit
3. Restriction Digestion: New England Biolabs (Beverly, MA) Manufacturer’s kit
4. Ligation: Rapid DNA Ligase (Roche Applied Science, Indianapolis, IN). Manufacturer’s kit
5. Chemical competent cell and Transformation (Heat shock) (Protocol developed by hands on experience)
6. PCR product purification kit and gel extraction kit (ZYMO RESEARCH)