Hi guys,
Just a note about PstI-HF:
The PstI-HF sold from NEB is less processive than the old NEB PstI/red stripe version (go here to see it: https://www.neb.com/products/r0140-psti) . When digesting ultrapure, high concentration homogeneous DNA (plasmids) digestions fail to go to completion even when left overnight (> 16 hours). What does this mean for you? It means that your genomes which are more difficult substrates to deal with will not digest fully unless left to go overnight. If you want to ensure a good digest use a positive control and switch to the linked product.