Digesting clones

Today I did a plasmid prep of some overnight E. coli transformants and set up a digestion. These clones were found to have the rec1 insert by PCR, I’m digesting with Hind III to linerize and mlul I to check directionality. Hind III is a relatively slow acting restriction enzyme (10,000 units needed to digest 1 ug of DNA in 1 hour) so I’m leaving them in the 37 degree incubator overnight. Tommorow I’ll run a gel and if all goes well I have a 87.5% chance of finding clones with the insert in both directions

 

Edit: My mistake! 10 000 units is the total amount of units you get when you purchase a tube from NEB. 1 unit is needed to digest 1 ug of DNA in 1 hour. I did not need to leave the digest overnight

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