Can I use In-Fusion Cloning to clone GC-rich DNA fragments?
Can I use In-Fusion Cloning to clone GC-rich DNA fragments?
Yes, but special consideration should be given to the homologous overlaps of adjacent DNA fragments. Since In-Fusion Cloning is based on the annealing of these overlaps, it is important to take the GC content into account for the 15-bp homology. We have no specific data showing variability of current In-Fusion Cloning master mix performance depending on the GC content of the 15-bp overlap. However, the following results were obtained using a previous version of the kit—In-Fusion Advantage:
-15-bp homologous overlaps with GC content of 20–40% had little or no effect on the In-Fusion Advantage cloning efficiency.
-15-bp homologous overlaps with GC content of 60–80% showed a reduced In-Fusion Advantage cloning efficiency in certain cases.In-Fusion Cloning products