The following is a list of 5' (back) primers that are based at the 5' end of FR1 of the VH, VK and VL segments. This is a miminal set which amplify all functional V genes and contain a maximum of a single degenerate position. An appropriate restriction site would need to be added to the 5' end for cloning purposes (see also restriction enzyme cuts in V genes).


VH primers

VH1a CAGGT(GT)CAGCTGGTGCAG VH1b CAGGTCCAGCTTGTGCAG VH1c (GC)AGGTCCAGCTGGTACAG VH1d CA(AG)ATGCAGCTGGTGCAG VH2a CAGATCACCTTGAAGGAG VH2b CAGGTCACCTTGA(AG)GGAG VH3a GA(AG)GTGCAGCTGGTGGAG VH3b CAGGTGCAGCTGGTGGAG VH3c GAGGTGCAGCTGTTGGAG VH4a CAG(CG)TGCAGCTGCAGGAG VH4b CAGGTGCAGCTACAGCAG VH5a GA(AG)GTGCAGCTGGTGCAG VH6a CAGGTACAGCTGCAGCAG VH7a CAGGT(CG)CAGCTGGTGCAA


VK primers

Vk1a (AG)ACATCCAGATGACCCAG Vk1b G(AC)CATCCAGTTGACCCAG Vk1c GCCATCC(AG)GATGACCCAG Vk1d GTCATCTGGATGACCCAG Vk2a GATATTGTGATGACCCAG Vk2b GAT(AG)TTGTGATGACTCAG Vk3a GAAATTGTGTTGAC(AG)CAG Vk3b GAAATAGTGATGACGCAG Vk3c GAAATTGTAATGACACAG Vk4a GACATCGTGATGACCCAG Vk5a GAAACGACACTCACGCAG Vk6a GAAATTGTGCTGACTCAG Vk6b GATGTTGTGATGACACAG


VL primers

VL1a CAGTCTGTGCTGACTCAG VL1b CAGTCTGTG(CT)TGACGCAG VL1c CAGTCTGTCGTGACGCAG VL2 CAGTCTGCCCTGACTCAG VL3a TCCTATG(AT)GCTGACTCAG VL3b TCCTATGAGCTGACACAG VL3c TCTTCTGAGCTGACTCAG VL3d TCCTATGAGCTGATGCAG VL4 CAGC(CT)TGTGCTGACTCAA VL5 CAG(CG)CTGTGCTGACTCAG VL6 AATTTTATGCTGACTCAG VL7 CAG(AG)CTGTGGTGACTCAG VL8 CAGACTGTGGTGACCCAG VL4/9 C(AT)GCCTGTGCTGACTCAG VL10 CAGGCAGGGCTGACTCAG