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To construct heavy and light linear expression cassettes by overlapping PCR, 10 ng of heavy or light chain fragments were used in a 50 L PCR reaction containing 25 L of Q5 hot start high fidelity DNA polymerase 2 Master Mix (New England Biolabs Cat # M0494L), 10 ng of the CMV fragment, 10 ng of the IgG or Kappa constant fragment, 20 pmol of the forward primer (gaccgagcgcagcgagtc) and 20 pmol of the reverse primer (tctccgagggatctcgacc)

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