EcoRI problem

Kris W. kwilde at uvic.ca
Thu Jul 10 11:09:56 EST 1997


The other day I was digesting a PCR-Script vector with EcoRI and I found
that it cut at two sites when I was only expecting it to cut at one.  I
normally leave the digestion for an hour or so but this time I left it
for 3 hours.  I re-did the digestion for a half hour and found that
there was only a single cut.  After looking at the sequence and the
sizes of the fractions caused by the digestion I concluded that EcoRI
(gaattc) cuts at aaattc when left for too long.  Similar sites such as
gaattg were not cut.  

Has anyone else noticed this or similar incidences with other
restriction enzymes.



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