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  • FAQ: What are the longest overlaps that can be used with this method?

    The quantity of 5´ exonuclease in the Gibson Assembly Master Mix and a 15 minute assembly reaction time have been optimized for the assembly of DNA molecules with ≤ 25-bp overlaps. If assembly reaction time is increased to 60 minutes, overlaps up to 40-bp may be used with the Gibson Assembly Cloning Kit.