Table 2 Primers used for generating Fab expressing phage display libraries. Degenerate nucleotides were used in few primers according to the IUPAC nomenclature (R = A/G, K = G/T, S = C/G, Y = C/T).
RT primers | |
IGHG1,2 A,2B,2 C | CTGGACAGGGATCCAGAGTTCCA |
IGHG3 | CTGGACAGGGCTCCATAGTTCCA |
IgK | GGGTGAAGTTGATGTCTTGTGAGTGGC |
PCR1 primers for variable-kappa chain amplification | |
VKappa1 | GCCACCATGGACATGAGG |
VKappa2 | GCCACCATGGGGTCCCAG |
VKappa3 | GCCACCATGGTGTCCCCGTT |
VKappa4 | GCCACCATGGAAACCCCA |
VKappa5 | GCCACCATGGTGTTGCAG |
VKappa6 | GCCACCATGGAAGCCCCA |
VKappa7 | GCCACCATGGACATGAGA |
VKappa8 | GCCACCATGGAACCATGG |
PCR1 primers for variable-heavy chain amplification | |
VH1 | GCCACCATGGACTGGACC |
VH2 | GCCACCATGGACTGCACC |
VH3 | GCCACCATGGACTGGATTTGGAGGG |
VH4 | GCCACCATGGARTTGGGG |
VH5 | GCCACCATGGAGTTTGGG |
VH6 | GCCACCATGGAGTTKGGACTGAGC |
VH7 | GCCACCATGGAGTTTTGGCT |
VH8 | GCCACCATGGAATTTGGGC |
VH9 | GCCACCATGGAACTGGGG |
PCR1 reverse primers are the same as used for RT reaction | |
PCR2 forward primers for variable-kappa chain amplification | |
ForIGKV1-6 | GCCATCCAGATGACCCAGTCTCC |
ForIGKV1-8 | GCCATCCGGATGACCCAGTCTCC |
ForIGKV1-17 | GACATCCAGATGACCCAGTCTCC |
ForIGKV2-24 | GATATTGTGATGACCCAGACTCC |
ForIGKV3-20 | GAAATTGTGTTGACGCAGTCTCC |
ForIGKV3D-7 | GAAATTGTAATGACACAGTCTCC |
ForIGKV4-1 | GACATCGTGATGACCCAGTCTCC |
ForIGKV6D-41 | GATGTTGTGATGACACAGTCTCC |
PCR2 forward primers for variable-heavy chain amplification | |
ForIGHV3-A | GARGTGCAGCTGKTGGAGTCTGG |
ForIGHV3-33 | CAGGTGCAGCTGGTGGAGTCTGG |
ForIGHV4-A | CAGSTGCAGCTGCAGGAGTCTGGCCCAGGACTG |
ForIGHV4-34 | CAGGTGCAGCTACAGCAGTGGGG |
ForIGHV6-1 | CAGGTACAGCTGCAGCAGTCAGG |
ForIGHV1-A | CARRTGCAGCTGGTGCAGTCTGG |
ForIGHV5-51 | GAGGTGCAGCTGGTGCAGTCTGG |
ForIGHV1-B | CAGGTYCAGCTGGTRCAGTCTGG |
PCR2 reverse primers for variable-kappa chain amplification | |
revKJ1 | TTTGATTTCCACCTTGGTCCC |
revKJ2 | TTTGATCTCCAGCTTGGTCCC |
revKJ3 | TTTGATATCCACTTTGGTCCCAG |
revKJ4 | TTTGATCTCCACCTTGGTCCC |
revKJ5 | TTTAATCTCCAGTCGTGTCCCTT |
PCR2 reverse primers for variable-heavy chain amplification | |
revHJ2 | TGAGGAGACAGTGACCAGGGT |
revHJ3 | TGAAGAGACGGTGACCATTGTC |
revHJ4 | TGAGGAGACGGTGACCAGG |
revHJ6 | TGAGGAGACGGTGACCGTG |
