Table 1.
Primers used for the generation of recombinant mouse monoclonal antibodies.
| Primers used for the generation of a leader sequence for the κ light chains | |
|---|---|
| 1 | GCCACCATGGAGTTTCAGACCCAGGTACTCATGTCCCTGCTGCTCTGCATGTCTGGTGC |
| 2 | GGCCGCACCAGACATGCAGAGCAGCAGGGACATGAGTACCTGGGTCTGAAACTCCATGGTGGC |
| Degenerate forward primers used for the amplification of rearranged κ light chains | |
| 3 | TCAATAGTTGAACATAGCGGCCGCASAAAWTGTKCTCACCCAGTC |
| 4 | TCAATAGTTGAACATAGCGGCCGCAGAWATTGTGCTMACTCAGTC |
| 5 | TCAATAGTTGAACATAGCGGCCGCAGACATTGTGCTRACACAGTC |
| 6 | TCAATAGTTGAACATAGCGGCCGCAGACATTGTGATGACMCAGTC |
| 7 | TCAATAGTTGAACATAGCGGCCGCAGAYATCMAGATRAMCCAGTC |
| 8 | TCAATAGTTGAACATAGCGGCCGCAGAYATCCAGATGAYTCAGTC |
| 9 | TCAATAGTTGAACATAGCGGCCGCAGATATCCAGATGACACAGAC |
| 10 | TCAATAGTTGAACATAGCGGCCGCAGACATTGTGCTGACCCAATC |
| 11 | TCAATAGTTGAACATAGCGGCCGCAGACATYSTRATGACCCARTC |
| 12 | TCAATAGTTGAACATAGCGGCCGCAGATRTTKTGATGACYCARAC |
| 13 | TCAATAGTTGAACATAGCGGCCGCAGAYATTGTGATGACBCAGKC |
| 14 | TCAATAGTTGAACATAGCGGCCGCAGATATTGTGATAACCCAGGA |
| 15 | TCAATAGTTGAACATAGCGGCCGCAGACATCYTGCTGACYCAGTC |
| 16 | TCAATAGTTGAACATAGCGGCCGCAGAAAWTGTGYTGACCCAGTC |
| 17 | TCAATAGTTGAACATAGCGGCCGCAGAAACAACTGTGACCCAGTC |
| 18 | TCAATAGTTGAACATAGCGGCCGCAGACATTRTGATGWCACAGTC |
| 19 | TCAATAGTTGAACATAGCGGCCGCAGACATCCAGMTGACMCARTC |
| Reverse primer used for the amplification of rearranged κ light chains | |
| 20 | GGATACAGTTGGTGCAGCATCAGCCC |
| Primers used for the generation of the joining fragment | |
| 21 | GGGCTGATGCTGCACCAACTGTA |
| 22 | TCAAGTGCAAAGACTCACTTTATTG |
| 23 | CAATAAAGTGAGTCTTTGCACTTGATAGTTATTAATAGTAATCAATTACG |
| 24 | TAGCAGAACAGGCAGCTTCATGGTGGCCTGGGAGTGGACACCTGTGGAGAG |
| 25 | ATGAAGCTGCCTGTTCTGCTAGTG |
| 26 | ACTGCTTGAGGCTGGACTCGTGAACAATAGCAGC |
| Degenerate forward primers used for the amplification of rearranged heavy chains | |
| 27 | TTCACGAGTCCAGCCTCAAGCAGTGAKRTRCAGCTTMAGGAGTC |
| 28 | TTCACGAGTCCAGCCTCAAGCAGTGAGKTYCAGCTBCAGCAGTC |
| 29 | TTCACGAGTCCAGCCTCAAGCAGTCAGGTGCAGMTGAAGSAGTC |
| 30 | TTCACGAGTCCAGCCTCAAGCAGTGAGRTCCAGCTGCAACARTC |
| 31 | TTCACGAGTCCAGCCTCAAGCAGTCAGGTYVAGCTGCAGCAGTC |
| 32 | TTCACGAGTCCAGCCTCAAGCAGTCAGGTYCARCTGCAGCAGTC |
| 33 | TTCACGAGTCCAGCCTCAAGCAGTGAGGTGMAGCTGGTGGAATC |
| 34 | TTCACGAGTCCAGCCTCAAGCAGTGAVGTGMWGCTSGTGGAGTC |
| 35 | TTCACGAGTCCAGCCTCAAGCAGTGARGTGCAGCTGKTGGAGWC |
| 36 | TTCACGAGTCCAGCCTCAAGCAGTGAGGTGAAGCTGATGGAATC |
| 37 | TTCACGAGTCCAGCCTCAAGCAGTGAGGTGCAGCTTGTTGAGTC |
| 38 | TTCACGAGTCCAGCCTCAAGCAGTGAGGTGAAGCTTCTCRAGTC |
| 39 | TTCACGAGTCCAGCCTCAAGCAGTGAAGTGAARMTTGAGGAGTC |
| 40 | TTCACGAGTCCAGCCTCAAGCAGTCAGGTTACTCWGAAAGWGTCTG |
| 41 | TTCACGAGTCCAGCCTCAAGCAGTCAGGTCCAACTGCAGCAGCC |
| 42 | TTCACGAGTCCAGCCTCAAGCAGTGATGTGAACCTGGAAGTGTC |
| 43 | TTCACGAGTCCAGCCTCAAGCAGTCAGATCCAGTTSGTRCAGTC |
| 44 | TTCACGAGTCCAGCCTCAAGCAGTGAGGTRCAGCTKGTAGAGAC |
| Reverse primers used for the amplification of rearranged heavy chains | |
| 45 | CTATTCTAGCTAATCTAGGCGCGCCGAGGAGACGGTGACCGTGGTCC |
| 46 | CTATTCTAGCTAATCTAGGCGCGCCGAGGAGACTGTGAGAGTGGTGC |
| 47 | CTATTCTAGCTAATCTAGGCGCGCCGCAGAGACAGTGACCAGAGTCC |
| 48 | CTATTCTAGCTAATCTAGGCGCGCCGAGGAGACGGTGACTGAGGTTC |
| Primers used for the amplification of the IgG1 constant region | |
| 49 | ATATCACGGCGCGCCCGACACCCCCATCTGTCTATCCA |
| 50 | GACTGATCTAGATCATTTACCAGGAGAGTGGGAGAG |
| Primers used for fusion polymerase chain reaction | |
| 51 | TCAATAGTTGAACATAGCGGCCGC |
| 52 | CTATTCTAGCTAATCTAGGCGCGCCG |
| Primer specific for the aberrant κ light chain from SP2/0 myeloma | |
| 53 | GGATGCTGCAACCTATTACTGTCAGCACATTAGGGAGCTTACACG |
The primers used to amplify the rearranged heavy and kappa light variable regions were designed using recent genome sequence of the mouse immunoglobulin regions: NG_005612 for the κ light locus and NG_005838 for the heavy locus. Degenerate base code: M = A+C; R = A+G; W = A+T; S = C+G; Y = C+T; K = G+T; V = A+C+G; B = C+G+T.