PCR Amplification and Plasmid Isolation
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Corresponding Organization :
Other organizations : Delft University of Technology, DSM (Netherlands), Amyris (United States)
Protocol cited in 1 other protocol
Variable analysis
- PCR amplification method (Phusion Hot Start II high-fidelity polymerase)
- Oligonucleotide primer purification method (HPLC or PAGE)
- DNA fragment size and quality (as observed on agarose gels)
- Plasmid isolation efficiency (from E. coli and yeast)
- E. coli transformation efficiency (chemical or electroporation)
- Manufacturer's instructions for Phusion Hot Start II polymerase, oligonucleotide primers, plasmid isolation kits, and transformation methods
- Use of standard Tris-acetate-EDTA buffer for agarose gel electrophoresis
- Use of DreamTaq polymerase for diagnostic PCR
- Use of E. coli DH5α strain for transformation
- None specified
- None specified
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