Primer

Part:BBa_K4647021:Design

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


IVA primer designed to anneal to nifD and amplify with homology to 2BCT


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 nifD 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

IVA primer designed to anneal to nifD and amplify with homology to 2BCT. This primer, with its FW pair, works to assemble the chunk nifB-nifH-nifD to the plasmid 2Bc-T.

References