Recipe

Isothermal Reaction (Gibson Assembly) Master Mix

ISO buffer (5×)   40 µL
T5 exonuclease (Epicentre T5E4111K)  1.6 µL
Phusion high-fidelity DNA polymerase (New England BioLabs F-530L)  2.5 µL
Taq ligase (New England BioLabs M0208L)   20 µL
H2O 85.9 µL
Total volume  150 µL
Combine the above ingredients. Store 15-µL aliquots at −20°C. Do not use a hot start polymerase.
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  1. doi:10.1101/pdb.rec090019 Cold Spring Harb Protoc 2017: pdb.rec090019- © 2017 Cold Spring Harbor Laboratory Press
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