Difference between revisions of "Part:BBa K4769000:Design"
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| + | ===Design Notes=== | ||
| + | pMiniMAD contains an internal BsaI cut site within the ampicillin resistance gene – this can be avoided when using BsaI by ensuring minimal homology of the internal 4 bp overhang with other designed overhangs and performing an additional final ligation step following Golden Gate assembly. | ||
| + | Alternatively, the BsaI recognition sequence could be removed by point mutation to a synonymous codon, by site-directed mutagenesis or using an additional internal set of primers. | ||
Latest revision as of 15:02, 8 October 2023
Design Notes
pMiniMAD contains an internal BsaI cut site within the ampicillin resistance gene – this can be avoided when using BsaI by ensuring minimal homology of the internal 4 bp overhang with other designed overhangs and performing an additional final ligation step following Golden Gate assembly.
Alternatively, the BsaI recognition sequence could be removed by point mutation to a synonymous codon, by site-directed mutagenesis or using an additional internal set of primers.