Table 2.
Sequence of the primers used to amplify and fuse the kappa light variable chain genes into an scFv phage display library.
| VLK family | Primer | Nucleotide sequence |
|---|---|---|
| Family 1 | MP20913 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGATGTTGTG + ATGACACAAACTCC |
| MP20914 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGATGTTGTGTTG + ACACAAACTC | |
| MP20915 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGATGTTGTGATGACCCAGACACCAC | |
| MP20916 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGATATTGTG + ATGACMCAGACTCC | |
| MP20917 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGATGTTGTGMTGACCCAGACTCCA | |
| Family 2 | MP20918 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGATAT + TGTGATGACTCAAGCTCC |
| MP20919 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGATATTGTGATGACCCAGGGTGCAC | |
| MP20920 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGATATCATGATGACTCAGTCTCCCCTCTC | |
| Family 3 | MP20921 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATTGTGCTGACCCAGTCTCC |
| MP20922 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATTGTCTTGACCCAGTCTCCTG | |
| MP20923 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACACTGTRCTGACCCAGTCTCC | |
| Family 4 | MP20924 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGAAATTGTGCTCACTCAGTCTCCAACAAC |
| MP20925 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGAA + ATTGTGCTCAYCCAGTCTC | |
| MP20926 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGATAATGTGCTCCCTCAGTCTCCAAC | |
| MP20927 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGAAACTGTGCTCACTCAATCTCCAACC | |
| MP20928 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGCTATTGTTCTCAACCAGTCTCCATCCATC | |
| MP20929 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGAAAT + TATACTCACCCAGTCTCC | |
| Family 5 | MP20930 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCGTGCTRACTCAGTCTCCA |
| MP20931 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACRTTGTGYTGACTCAGTCTCCAGC | |
| Family 6 | MP20932 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACAYTGTGATGACCCAGTCTCC |
| MP20933 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGAACAYTGTGATGACYCAGTCTCCCA | |
| Family 7 | MP20934 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGAATGTCATGATGACCCAGTCTCCAAC |
| Family 8 | MP20935 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACAT + TGTGATRACCCARTCT |
| MP20936 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATTGCGATAACCCAGTCTCC | |
| MP20937 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACAT + TKTGATGAC + CCAG | |
| MP20938 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATTTTGATAAACCAGTCTCCAGCCTC | |
| Family 9 | MP20939 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGCAGATCACGCTCACCCAGCAAG |
| Family 10 | MP20940 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGWTGACCCAGTCTCCATC |
| MP20941 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATC + CAGWTKACCCAG | |
| Family 12 | MP20942 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCA + GATGACACAGTCTCC |
| MP20943 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGATATCCRGATG + ACWCAGTCTCC | |
| Family 14 | MP20944 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCAGATGACCCAGTCTCC |
| MP20945 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATTCAGATGACCCAGTCTCCATCC | |
| MP20946 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATTCAGATGACSCAGKCTYCATC | |
| Family 15 to 20 | MP20947 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGATRTCCAGATG + ACMCAGTC |
| MP20948 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGAAACTACTGTGACCCAGTCTCCAGC | |
| MP20949 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGACTGGAGAAACAACACAGTCTCCAGC | |
| MP20950 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATTAGGATGACTCAGACTCCAGC | |
| MP20951 | GGCGGAGGTGGCTCTGGCGGTGGCGGATCGGACATCCACATGACTCAGAACCCAG | |
| Joining region (KJ) | MP23454 | TGCACGCTGCTAGATATGAGGCACTGCGGCCGCGTTTCAATTCCAGCTTGGTGCCTCC |
| MP23455 | TGCACGCTGCTAGATATGAGGCACTGCGGCCGCGTTTCAGTTCCAGCTTGGTCCCAG | |
| MP23456 | TGCACGCTGCTAGATATGAGGCACTGCGGCCGCGTTTTATTTCCAGTCTGGTCTCATCACTG | |
| MP23457 | TGCACGCTGCTAGATATGAGGCACTGCGGCCGCGTTTTATTTCCAACTTCGTCCCTGAGCC | |
| MP23458 | TGCACGCTGCTAGATATGAGGCACTGCGGCCGCGTTTGATCTCCAGCTTGGTCCCAGAAC | |
| Overlap extension PCR | VLK outer primer | TGCACGCTGCTAGATATGAGGCAC |
The table includes forward and reverse primer specific for different genes of the kappa light chain families and joining regions of Rattus norvegicus germline sequences. An outer reverse primer was used for the overlap extension PCR for the assembly as scFv phagemid library. The primer may include an ambiguous DNA nucleotide (Y, R, W, S, K and M) and Locked Nucleic Acids (LNAs) were incorporated in specific conserved residues of the variable chains’ genes facilitating the generation of short primers to avoid extension into the frame, minimizing mismatches in other regions of the primer. Five reverse primer were designed to cover all the joining regions of the VLK.