TABLE 1.
Oligonucleotide primers used for construction of the library and sequencing
| Purpose | Primer name | Sequencea |
|---|---|---|
| VH | VH1back (a) | 5′aataggcccagccggccatggcc(C/G)AGGTTCA(A/G)(G/C)TGCAGCAG(C/T)CTGG3′ |
| VH2back (a) | 5′aataggcccagccggccatggccGAGGTG(C/A)AGCTT(G/C)T(G/C)GAGTCTGG3′ | |
| VH3back (a) | 5′aataggcccagccggccatggccCCGA(G/T)GTGCAGCTTCAGGAGTCAGG3′ | |
| VHfor | 5′CCTGAGGAGACGGTGACC(A/G/C)(G/T)GG(T/G)(T/C)CC3′ | |
| IgG1-CHfor | 5′TGGATAGACAGATGGGGG3′ | |
| Vκ | VK1back | 5′ggatctGACATC(C/A)AGATGACTCAG(T/A)CT3′ |
| VK2back | 5′ggatctGATATTGTGATGACGCAG(G/A)(C/A)T3′ | |
| VK3back | 5′ggatct(G/A)ACATTGTG(C/A)TGTCACAGTCTCC3′ | |
| VK4back | 5′ggatctGAAA(A/T)T(G/T)TTCTCACCCAGTCTCC3′ | |
| VK5back | 5′ggatctGATATT(G/T)TGATGACGCAG(C/T)CTGC3′ | |
| VKfor (a) | 5′aatggaattccgggcggccgcCCGTTTTA(T/G)(T/C)TCCAGCTT(G/T)GT(G/C)CC3′ | |
| CKfor | 5′AGTTGGTGCAGCATCAGCC3′ | |
| Linker | VHLINKback | 5′GGTCACCGTCTCCTCAgg3′ |
| VK1LINKfor | 5′AG(A/T)CTGAGTCATCT(G/T)GATGTCagatcc3′ | |
| VK2LINKfor | 5′CTGCGTCATCACAATATCagatcc3′ | |
| VK3LINKfor | 5′GGAGACTGTGACA(G/T)CACAATGT(C/T)agatcc3′ | |
| VK4LINKfor | 5′GGAGACTGGGTGAGAA(C/A)A(T/A)TTTCAGATagatcc3′ | |
| VK5LINKfor | 5′GCAG(G/A)CTGCGTCATCA(C/A)AATATCagatcc3′ | |
| Sequencing | geneIII | 5′TGAATTTTCTGTATGAGG3′ |
| linkfwd | 5′CTCGGGCGGTGGTGGGTC3′ | |
| linkrev | 5′GACCCACCACCGCCCGAG3′ |
a
The SfiI sites in the VHback primer sequences and the NotI site in the VKfor primer sequence are underlined. Nucleotides represented as uppercase characters are carried by the V gene; those in lowercase characters are carried by the vector.