Primer
Part:BBa_K4436017:Design
Designed by: Vedanta Kompella Group: iGEM22_UIUC_Illinois (2022-10-11)
Forward Primer for Lambda Red CsgA Knockout
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
We had to ensure the GC content of the primers was low enough that the annealing temperature for Q5 PCR wasn't too high.
Source
It has 40 nucleotide homology to the flanking regions of the csgA gene, and 20 nucleotide homology to the flanking regions of the FLP recombinase sites in pKD3.