Table 2.
Nucleotide sequence of inverse PCR primers.
| Gene | Primer sequence 5′ to 3′ | Co-ordinatesa | Restriction enzymeb |
|---|---|---|---|
| ligA 5′ end | CAGATATTCTTACCGTTTCCAATACA | 1883–1908 | NsiI |
| ATCCGATCAGATTTTTGCATCAAGTC | −209–(−234) | ||
| ligA 3′ end | CCGCTTTCTGTAGGTTCCTCTAAAAT | 3562–3587 | HincII |
| CGGTAGCGGTCAGTTGTAGTGTAAGA | 3175–3150 | ||
| ligB 3′ end | ATATCCGGAATGAATTTTGGTGTAAA | 5569–5594 | EcoRI |
| ATTTTCAAGATTTGTTCTCCAGATTT | 5487–5462 | ||
| LigC 5′ end | TCCTGCAAATCCAAGCGTAGCCAATG | 1054–1079 | XhoI |
| GTAAGAACTGCGGAAGTGACGTTTACTA | 1038–1011 | ||
| LigC 3′ end | ACCGGACTCTTGTTCTAACTGGGTGG | 5209–5234 | BglII |
| ATCCGATTCCGTCGTTGCTTGACTTT | 4868–4843 |
a
Relative to A in ATG start codon.
b
Leptospiral DNA digested with restriction enzyme before treatment with T4 ligase and PCR.