I have a vector with 5308bp length.
I digested this vector by NotI and ECORI enzymes. I must be observe two bands 1346bp and 3962bp but I observed 1346bp and 2500bp. What I observed band at 2500bp instead of 3962bp?
I ran digestion product on gel 0.6℅.
Hi there,
With a picture comparing uncut and cut plasmids, it would be better.
Are you sure of the starting plasmid and of your gel calibration?
I would advise to run double cut and single enzyme cut vector running at same gel. With each single enzyme cut you should see a band of 5308. If you see a band of 3846 bp, it will mean that vector has a problem.
Hi there,
When I took a look at the sides of your bands, I could see that 3962 bp minus 1346 bp is equal to 2616 bp.
That is quite close to 2500 bp. So I personally think that EcoR1 or Not 1 is cutting your vector more than twice, Leading to two DNA fragments with a length close to 1346 bp for each, and a third fragment close to 2500 bp.
Check your templates for the different EcoR1 and Not1 sites on your plasmid.
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