TABLE 1.
Signature tag sequences and primers
| Oligonucleotidea | Sequence (5′→3′) |
|---|---|
| STM-1L-fwd | GGCCGCCCGGGTTAAACGTTCAACTTGCAGGGCC |
| STM-1L-rev | CTGCAAGTTGAACGTTTAACCCGGGC |
| STM-1L-Tag | GGGCCCTGCAAGTTGAACG |
| STM-2L-fwd | GGCCGCCCGGGTTAAATAGCCTATAGCCTGGGCC |
| STM-2L-rev | CAGGCTATAGGCTATTTAACCCGGGC |
| STM-2L-Tag | GAGGGCCCAGGCTATAGGC |
| STM-3L-fwd | GGCCGCCCGGGTTAAAAGTCTCAAGTCTCGGGCC |
| STM-3L-rev | CGAGACTTGAGACTTTTAACCCGGGC |
| STM-3L-Tag | GGGCCCGAGACTTGAGAC |
| STM-4L-fwd | GGCCGCCCGGGTTAATAACGTGTAACGTGGGGCC |
| STM-4L-rev | CCACGTTACACGTTATTAACCCGGGC |
| STM-4L-Tag | GAGGGCCCCACGTTACACG |
| STM-5L-fwd | GGCCGCCCGGGTTAAACTGGTAACTGGTAGGGCC |
| STM-5L-rev | CTACCAGTTACCAGTTTAACCCGGGC |
| STM-5L-Tag | GGGCCCTACCAGTTACCAG |
| STM-6L-fwd | GGCCGCCCGGGTTAAGCATGTTGCATGTTGGGCC |
| STM-6L-rev | CAACATGCAACATGCTTAACCCGGGC |
| STM-6L-Tag | GGGCCCAACATGCAACATG |
| STM-7L-fwd | GGCCGCCCGGGTTAATGTAACCCCAATGTGGGCC |
| STM-7L-rev | CACATTGGGGTTACATTAACCCGGGC |
| STM-7L-Tag | GGGCCCACATTGGGGTTAC |
| STM-8L-fwd | GGCCGCCCGGGTTAAAATCTCGGCTCTAAGGGCC |
| STM-8L-rev | CTTAGAGCCGAGATTTTAACCCGGGC |
| STM-8L-Tag | GAGGGCCCTTAGAGCCGAG |
| STM-9L-fwd | GGCCGCCCGGGTTAATAGGCAAAACGGATGGGCC |
| STM-9L-rev | CATCCGTTTTGCCTATTAACCCGGGC |
| STM-9L-Tag | GGGCCCATCCGTTTTGCC |
| STM-10L-fwd | GGCCGCCCGGGTTAACAATCGTTGCTAACGGGCC |
| STM-10L-rev | CGTTAGCAACGATTGTTAACCCGGGC |
| STM-10L-Tag | GAGGGCCCGTTAGCAACG |
| STM-11L-fwd | GGCCGCCCGGGTTAATCAAGACCAGAACTGGGCC |
| STM-11L-rev | CAGTTCTGGTCTTGATTAACCCGGGC |
| STM-11L-Tag | GGGCCCAGTTCTGGTCTTG |
| STM-12L-fwd | GGCCGCCCGGGTTAACTAGTAGGATGATCGGGCC |
| STM-12L-rev | CGATCATCCTACTAGTTAACCCGGGC |
| STM-12L-Tag | GAGGGCCCGATCATCCTAC |
a
Forward (fwd) and reverse (rev) oligonucleotides were annealed to form the tag that was inserted between the NotI and ApaI sites on pJDM-STM vectors. STM-#L-Tag refers to the specific primer used in the PCR mixtures to determine the presence of a specific mutant clone in the pool.