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Tag Archives: Golden Gate Cloning

Golden Gate Cloning

Materials needed: BsaI, NEB# R0535, Antarctic Phosphatase, NEB#M0201 Polynucleotide kinase, xxx T4 ligase T4 ligase buffer Mach 1 chemically competent cells HF BsaI HF BamHI Procedure: Digest and dephosphorylate plasmid. Use BsaI (NEB# R0535, standard enzyme), NEB buffer 4. Digest with 1 ul of enzyme for 1 hr. 37°C Add 1 ul more of enzyme…

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