Back to Lidstrom:Protocols
Janet is developing a protocol here
- overlap by 40 bp
Oligo (primer) Design
- Electroporation can give much higher efficiency. For this reason, the assembled product is usually electroporated.
- rxn product is salty: can be a problem for electrocompetent cells. Rob Egbert dilutes the competent cells when transforming with electroporation. Not so much a problem when using chemically competent cells.
- Janet electroporates 2 uL
- Colony PCR 1st
- Sequence a few clones with primers that cover the "seams."
- Electrocompetence is ~ 1000x more effective so is tempting. -