Primer

Part:BBa_K3406008:Design

Designed by: Xiaojie Zhou   Group: iGEM20_ZJUT_China_B   (2020-10-27)
Revision as of 07:14, 27 October 2020 by Sorrow-filter (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

RPA forward primer for H1N1


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 31
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 31
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 31
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We use Recombinase Polymerase Amplification to amplify a short sequence of H1N1 genome. To maximize amplification success, the primer lengths should be ~25–35 nt, and the total amplicon size should be 80–140 bp. Primers are typically designed with melting temperatures between 54 and 67 °C. In addition, because we need to do transcription experiments downstream, a T7RNA polymerase promoter was appended to the 5ʹend of the forward primers to allow T7 transcription.

Source

References