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:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE 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.

References