Ligation Protocol using BioBrick Assembly Kit® (E0546)


Ligate the Upstream and Downstream Parts into the digested Destination Plasmid.

Upstream Part digestion: 2 µl
Downstream Part digestion: 2 µl
Destination Plasmid digestion: 2 µl
10X T4 DNA Ligase Buffer: 2 µl
T4 DNA Ligase: 1 µl
H2O: 11 µl

Incubate at room temperature for 10 minutes and then heat inactivate at 80°C for 20 minutes.

Transform 2 μl of the ligation product into 50 μl of competent E. coli cells (or other suitable host strain). Select using the antibiotic corresponding to the Destination Plasmid. 

Please refer to the individual datacards for each reagent for recommended use and storage conditions. 

Technical questions regarding the individual use of any of the enzymes included in this kit should be directed to New England Biolabs by contacting 1-800-632-7799 or

Technical questions regarding the assembly of BioBrick® parts should be directed to Ginkgo BioWorks by emailing