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NEBuilder Assembly Tool can be used to design primers for your NEBuilder HiFi DNA or Gibson Assembly reactions, based on the entered fragment sequences and the polymerase being used for amplification.

Required Data:

To get started, please have the sequences of your DNA fragments ready in raw or FASTA format. NEBuilder Assembly Tool has many common vector sequences available for you, as well as a few sample insert sequences if you just want to learn. You should also have ready either the restriction sites or sequence coordinates that will define the insertion site on the vector.

Basic Usage:
  1. Set preferences for the assembly, including the number of fragments to be joined and the enzyme you will use for PCR amplification. Change settings at any time and the results will be instantly updated.
  2. Build your construct by adding fragments to your assembly. You can add one vector backbone and one or more inserts. Repeat as needed.
  3. View primers. NEBuilder Assembly Toolwill give you a list of the required primers, their melting temperatures, recommended annealing temperatures, as well as the final sequence of your assembled construct and a summary of the design.
Additional Information:

Set Preferences

Current preferences help tip

Build Construct

Fragment arrangement help tip
  • Click and drag insert fragments to rearrange their order. Double-click fragments or vectors to edit properties.
    Required Primers help tip