Restriction enzymes identify a specific recognition sequence in the DNA, and generate double-stranded
breaks in the DNA duplex. Cleavage results in fragments with either a 5' or 3' overhang,
commonly referred to as sticky ends, or no overhang, referred to as blunt ends.
The 5' phosphates and 3' hydroxyls are maintained after cleavage, leaving the DNA ends ready
to be joined via ligation. The ability of some restriction enzymes to
predictably cleave DNA and generate distinct DNA bands with ligatable ends has made them
an invaluable tool for recombinant DNA technologies. When selecting restriction enzymes for use
in a cloning experiment, it is important to: determine which enzymes will produce ends
compatible with the selected vector confirm that recognition sites do not occur
within the DNA fragment to be cloned and examine the methylation sensitivity
of your selected enzyme to confirm that host methylation by dam and/or dcm methylases will
not block cleavage. NEB offers a variety of online interactive tools to aid in selecting
which enzyme to use, as well as parameters for setting up reactions.
Visit CLONEWITHNEB.com for a full list of products available for this application.