The 1-base overhangs produced by AhdI may be hard to ligate. Sticky ends from different AhdI sites may not be compatible.
PciI (4983) 1 site
ACATGTTGTACA
PciI is inhibited by nonionic detergents.
BspQI (4867) 1 site
GCTCTTCNCGAGAAGNNNN
Sticky ends from different BspQI sites may not be compatible.
SapI (4867) 1 site
GCTCTTCNCGAGAAGNNNN
Sticky ends from different SapI sites may not be compatible.SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.