Primer

Part:BBa_K4479012:Design

Designed by: Alston Lo, Ruochen Liu, Xinyi Xuan   Group: iGEM22_Toronto   (2022-10-12)


F3 LAMP Primer for Oak Wilt 4


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 ranked our F3 primer based on the following criteria: 1. Distance between F2 and F3 primer: 0 - 60 nt 2. Length of F3 primer: 15 - 25 bp 3. GC Content: 40 - 60% 4. Melting temperature for F3 primer: 55 - 63℃ 5. Melting temperature difference between F3 and B3 primers: ≤ 5℃

Other criteria for extra marks: 1. No runs of 3 or more of one base (i.e ACCC) 2. No dinucleotide repeats (i.e ATATATAT) 3. Avoid forward and reverse primers having complementary sequences 4. Avoid more than three bases that complement within the primer



Source

The part was designed with the GLAPD software and then it was ranked against primers provided by the Primer Explorer software using computer models by our Drylab team. The criteria for ranking were: Primer Distance, Primer Length, GC Content, Melting Temperature, and Melting Temperature Difference. The target sequence it originated from is MG270170.1.

References