This can happen alot with cloning due to strange recombination events during lag phase of growth of transformants

The only advice I can give when you see this is pick 12 colonies miniprep and digest and hopefully 1 will be the correct recombinant

If none yield the correct insert then clone again

Also make sure your restriction digestion is complete if necessary by digesting for a minimum of 2 hours

Alternatively as a screen you could PCR inserts in the miniprep plasmids. This is often favoured today over restriction digestion as a mechanism of identifying correct recombinants

Thank you for your thoughts. I only picked two so I might try more colonies.

also, I did the digestion for 6 hours at 37C so I think that’s not the problem.

Finally, would try your recommendation (PCR inserts in the miniprep plasmid.