Primer

Part:BBa_K4647016:Design

Designed by: Bushra Islam   Group: iGEM23_Stony-Brook   (2023-10-12)


IVA primer designed to anneal to hesA and amplify with homology to nifX


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

The primer pairs to add the homology regions to the ends of hesA required a number of considerations. First, the annealing temperature to the template binding region had to be ~60C. The base pair count was kept around 20 base pairs. The homology regions had an annealing temperature of around 47-52C and had a base pair count of 15-20. Deviations from this design had either lowered efficiency or CFU.


Source

This primer was designed using our understanding of the 2016 in-vivo assembly paper (Garcia-Nafria 2016).

References