Table 2 Sequence of the primers used to amplify and fuse the kappa light variable chain genes into an scFv phage display library.

From: A primer set for the rapid isolation of scFv fragments against cell surface antigens from immunised rats

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

  1. 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.
Back to article page