Primer design is not one-size-fits-all. There are critical application-specific parameters to consider that can vastly increase your likelihood of experimental success. Whether you are creating an assembly that incorporates multiple fragments, conducting your entire PCR at a single temperature, or incorporating intentionally mismatched nucleotides, careful primer design can set you up

Here at NEB, we have created a variety of interactive tools to help you accurately design primers to suit your specific needs.

 

NEBuilder HiFi DNA Assembly
& Gibson Assembly

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NEBuilder® Assembly Tool

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

Loop-mediated Amplification
(LAMP)

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NEB LAMP Primer Design Tool

NEB LAMP Primer Design Tool can be used to design primers for your Loop-mediated Isothermal Amplification. Fixed primers can be specified for the design of LAMP primers, and subsequent Loop primers are then designed based on LAMP primer selection.

Golden Gate Assembly

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NEB Golden Gate Assembly Tool 

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

Site-directed Mutagenesis

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NEBaseChanger® 

NEBaseChanger can be used to design primers specific to the mutagenesis experiment you are performing using the Q5 Site-Directed Mutagenesis Kit. This tool will also calculate a recommended custom annealing temperature based on the sequence of the primers by taking into account any mismatches.

 

 

 

 

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