TABLE 1.
Oligonucleotides used for PCR
| Oligonucleotide | Positiona | Strand | Length (bases) | Sequence, 5′−3′b |
|---|---|---|---|---|
| Kan | NA | NA | 21 | CCTCTTCCGACCATCAAGCAT |
| 59F1 | 318–340 | + | 22 | GGCTAATAACCGGCGGAGAAAC |
| 59R1 | 5632–5612 | − | 20 | CCGTAAGTCGCTACATACTG |
| 59R2c | 5223–5243 | + | 20 | TTCTGCTGCTCTCTCTCAAG |
| Urease/Xba | 11120–11100 | − | 32 | CCGGCCTCTAGAAGCGAAGCACGAATTAAGTT |
| Urease/Sac | 4527–4548 | + | 32 | GCGGCGAGCTCAGTTGGTTATTATTGGCGGTT |
| Tn10 | NA | NA | 21 | GATCATATGACAAGATGTGTA |
a
Position 5′ to 3′ within the compiled urease and upstream sequences (see Fig. 1). Note that the Kan and Tn10 primers were designed from sites within the Kan cassette of and near the end of the mini-Tn10 transposon sequence, respectively. NA, not applicable.
b
Extensions containing the restriction enzyme-cut sites (in bold) are shown.