New England Biolabs    
Double Digest Finder (Utilizing old NEB Restriction Enzyme Buffer System)
Cleaving a DNA substrate with two restriction enzymes simultaneously (double digestion) is a common timesaving procedure. Use this tool to select reaction conditions amenable to any two NEB restriction enzymes.
Please note that as of October 1, 2015, this version of the Double Digest Finder will no longer be available. The transition to our CutSmart buffer system two years ago allowed us to simplify the double digest process and create an updated version of the tool.
Refer to the previous version of our Restriction Enzyme Activity Chart for guidance on how to use your old buffers in conjunction with the recommendations from the new tool. If you have any questions, please contact our technical support team at
We also encourage you to use our online tool, NEBcloner, which provides you with a detailed double-digest protocol based upon the 2 restriction enzymes that you select.
To perform a double digest calculation, please select both 1st and 2nd enzymes.
Select 1st enzyme:       Select 2nd enzyme:       

Click to access the new Double Digest Finder (Utilizing new NEB Restriction Enzyme Buffer System)