Primer

Part:BBa_K3839034

Designed by: Georgios Psarias   Group: iGEM21_Patras   (2021-10-21)

CYB5A_202 TOPO site Forward Primer

The Forward primers of each gene of interest are designed with a TOPO site (CACC) at the 5’ end of their sequence for directional cloning the blunt-end PCR product into a pENTR/D-TOPO vector. For this cloning procedure no ligase was used, or any restriction enzyme. In this system, PCR products are directionally cloned by adding four bases to the forward primer (CACC). The overhang in the cloning vector (GTGG) invades the 5′ end of the PCR product, anneals to the added bases, and stabilizes the PCR product in the correct orientation. Inserts can be cloned in the correct orientation with efficiencies equal to or greater than 90%.



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]
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Parameters
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