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amplification of Fab expression cassette...

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  1. cDNA is quantified by UV spectrometry as below.
  2. The rabbit VH and VK/VL genes are amplified from cDNA pool by the first round of PCR
  3. Both human CK and human CH1 genes are amplified from pDisplay-2 by the 2nd round of PCR.
  4. Rabbit VK/VL genes are assembled with human CK by conventional overlapping PCR. The product is named as light chain cassette.
  5. Rabbit VH genes are assembled with human CH1 by conventional overlapping PCR. The product is named as heavy chain cassette.
  6. The light chain cassette and the heavy chain cassette are assembled into Fab casstte by over-lapping PCR too.

The primers and the schematic process above are shown in appendices section.


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