Recommended Screening Protocols for Using NEB® Golden Gate Assembly Kit (BsaI-HF®v2)
- Colony PCR screening using an appropriate DNA polymerase for amplification of the insert region. While many DNA polymerases are suitable for colony PCR, OneTaq ® DNA Polymerase (NEB #M0480) or OneTaq Hot Start DNA Polymerase (NEB #M0481) are recommended. Taq DNA Polymerase (NEB #M0267) can also be used. For PCR of larger assemblies, LongAmp® Taq DNA Polymerase (NEB #M0323) or LongAmp Hot Start Taq DNA Polymerase (NEB #M0534) are strongly recommended.
- Preparing plasmid mini-preps using the Monarch Plasmid Miniprep Kit (NEB #T1010 ) and mapping by using appropriate restriction endonucleases to confirm the correct assembly.