Primers for the Construction of Marmoset Phage-Displayed Fab Libraries
- H chain variable region (IGHV) 5' primers:
CjFabVH1/7 5’-gctgcccaaccagccatggcccaggtgcagctggtgcagtctgg-3’
CjFabVH3/5 5’-gctgcccaaccagccatggccgaggtgcagctggtgsagtctgg-3’
CjFabVH4 5’-gctgcccaaccagccatggcccaggtgcagctgcaggagtcgga-3’
CjFabVH6 5’-gctgcccaaccagccatggcccaggtgcagctgcaggagtcagg-3’
- H chain variable region (IGHV) 3' primer:
CjCH1-B 5’-gcagggtgtcagggggaagacatctgggttcttggtggaggctga-3’
- H chain g constant region (IGCG-CH1) 5' primer:
CjCH1-F 5’-gtcttccccctgacaccctgcggc-3’
- H chain g constant region (IGCG-CH1) 3' primer:
gIIIA-R 5’-taatcaaaatcaccggaaccagag-3’
- H chain assembly
5' primer Lead-VH 5’-gctgcccaaccagccatggcc-3’
3’ primer gIIIA-R see above
- kL chain variable region (IGKV) 5' primers (SfiI site shaded in gray) (20):
HFabK1 5’-gggcccaggcggccgagctccagatgacccagtctcc-3’
HFabK2/4 5’-gggcccaggcggccgagctcgtgatgacycagtctcc-3’
HFabK3 5’-gggcccaggcggccgagctcgtgwtgacrcagtctcc-3’
HFabK5 5’-gggcccaggcggccgagctcacactcacgcagtctcc-3’
- kL chain variable region (IGKV) 3' primer:
CjCK-B 5’-gaagacagacggcgcagccacagc-3’
- kL chain constant region (IGKC) 5' primer:
CjCK-F 5’-cgagctgtggctgcgccgtctgtc-3’
- kL chain constant region (IGKC) 3' primer
Lead-B 5’-ggccatggctggttgggcagc-3’
- kL chain assembly (SfiI site shaded in gray):
5' primer RSC-F 5’-gaggaggaggaggaggaggcggggcccaggcggccgagctc-3’
3' primer Lead-B see above
- Assembly of full length Fab cloning product
5’ primer RSC-F see above
3’ primer gIIIB-R 5’-cagagccgccaccctcagaac-3’
PCR steps involved:
