NEBcutter V3.0

Use NEBcutter2.0 tool to find the restriction sites within your DNA sequence, identifiying the sites for both Type II and comercially available Type III restriction enzymes. Enter your DNA sequence (maximun length 300KBases) and click on “submit” to find the restriction sites.


NEBcutter V3.0 at



NEBcutter is a web-based tool for identifying restriction enzyme recognition sites in a DNA sequence. It is provided by New England Biolabs (NEB), a leading manufacturer of enzymes for molecular biology research. Here is a complete tutorial for using NEBcutter:
  • Enter your DNA sequence in the “Sequence Input” box. You can also upload a file containing your sequence, or paste in a GenBank or GenBank-formatted file.
  • Select the enzyme(s) you want to use for cutting the sequence. You can select multiple enzymes by holding down the “Ctrl” key.
  • Select the type of output you want to receive (e.g. fragment sizes, fragment sequences)
  • Click the “Cut” button to run the analysis
  • The results will be displayed in a table, showing the location of recognition sites and the resulting fragments for each enzyme selected.
Some other features of NEBcutter are:

You can also use it to cut specific regions of the sequence by entering the start and stop positions in the “Region of Interest” field.

You can also use it to compare the restriction enzyme cutting patterns of multiple sequences by uploading or entering them in the “Sequence Comparison” field.

Leave a Comment

Your email address will not be published.