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  • FAQ: The number of colonies that don't contain an insert seems high, how can I tell if the Antarctic Phosphatase worked?

    The following controls can be used to diagnose this problem:
    1. Uncut vector: to check cell competency and antibiotic resistance.
    2. Vector cut and not ligated: to check for uncut vector, gel purification may be required to remove the last 0.1% of uncut vector.
    3. Vector cut and ligated: to check for intact ends and ligation conditions.
    4. Vector cut, phosphatased and ligated: to check for dephosphorylation, should be 3-5% of the vector cut and ligated.

    Ligation conditions should also be checked.