HomeFAQsIs there a difference in cutting close to the ends between NcoI-HF and NcoI?
FAQ: Is there a difference in cutting close to the ends between NcoI-HF and NcoI?
No. When tested on a series of five oligos (19-23 bases in length) containing the restriction site and an additional 1 to 5 A/T bases from the end, both enzymes cut the oligo with 2 base pairs from the end. When designing primers, NEB recommends adding 6 extra bases to ensure against experimental failure due to variation in substrate and reaction conditions.