I have a problem with the restriction enzyme XhoI and NotI form NEB.
I'm traying to cut 1ug of DNA (purified by Pure yield Midiprep system Kit from Promega) but I see partial digestion in the agarose gel.
I've tried to digest since 500 ng to 5 ug but the results are the same. I used low concentration enzyme (20 U/ul) to digest 500 ng and 4 ug and high concentration enzyme (100 U/ul) to digest 500 ng 1 ug 2 ug and 5 ug. And I don't know wat I'm doing wrong.
I'm using the next conditions:
5 ul Buffer NEB2
0.5 ul BSA (100X)
38 ul DNA (130 ng/ul)
4.5 ul Water
1 ul XhoI (100 U/ ul)
1 ul NotI-HF (100 U/ul)
Incubation at 37 °C for 2 hours.
Desactivation 65 °C for 15 min.