TABLE 2.
Oligonucleotide primer sequences based on the CDS of pBtoxis
| CDS | Orientation | Oligonucleotide primer (5′-3′) | Tm (°C)a | Amplicon size (bp) |
|---|---|---|---|---|
| pBt011 | Forward | GACAATTATCGAAGTTGAAAAGG | 62 | 430 |
| Reverse | ACTATTGAATCCTTCCGTTTCC | 62 | ||
| pBt014 | Forward | ACAGTAATCTATCTAAAATTGAGC | 62 | 278 |
| Reverse | TCTAAAGTTTCACGTAGCTTCC | 62 | ||
| pBt020 | Forward | GAAGAACAAATTTTAGAGAAAGG | 60 | 128 |
| Reverse | TAGTAGCTATATTTATTAAATATGG | 60 | ||
| pBt021 | Forward | CTGGGGCAAGGAAACTGCTA | 62 | 270 |
| Reverse | TTAGAGGGTTCCATTAATAGCG | 62 | ||
| pBt023 | Forward | AGATAGTTCTTTAGATACTTTAAG | 60 | 314 |
| Reverse | CATCACTAACCTCAATAATCC | 60 | ||
| pBt029 | Forward | TATTAGGTTCAGAAAAGAATCCG | 62 | 223 |
| Reverse | CAACAGAACTTATATTTCTTTCC | 60 | ||
| pBt031 | Forward | GGTGCACATGATTCAGGTGC | 62 | 679 |
| Reverse | CGATGTGCATTACCTTCAAGG | 62 | ||
| pBt036 | Forward | AGGCATATTGCATTAACAGTTCC | 64 | 411 |
| Reverse | TGTTTGAGTAGCTGATAAATTACG | 64 | ||
| pBt054 | Forward | GCACTAGTACCTACTACATCC | 62 | 703 |
| Reverse | GGTTTTGAATACTGTAAGCACG | 62 | ||
| pBt056 | Forward | GATGATTTGTTTAATCCTGAATGG | 64 | 323 |
| Reverse | TTCATTGTGGGGAAACTTTGC | 60 | ||
| pBt060 | Forward | GAATGAAAATATTGAAACCATCC | 60 | 391 |
| Reverse | AGAAATAGGTTCCTGAATCG | 60 | ||
| pBt063 | Forward | TTATTCATTCGCTACAACTTGC | 60 | 192 |
| Reverse | CAAAATAACTCGTTAGAATCTGG | 62 | ||
| pBt075 | Forward | AGGAGATAGTCTAAGATATGTACG | 66 | 324 |
| Reverse | GTATTACCATTTAGATCTGTGACG | 66 | ||
| pBt084 | Forward | TAATTGCCTTAGATAGAACACC | 60 | 541 |
| Reverse | GAGATTTTGGTTAAGTACTTTCC | 62 | ||
| pBt085 | Forward | TATTCACTCACTGCAACTGGC | 62 | 623 |
| Reverse | CTGTATTCGCTATCAGTATTCC | 62 | ||
| pBt086 | Forward | GAATGAGATTGTTGAAACTATCC | 62 | 641 |
| Reverse | AGTTAGATAATTCTCCTATGCC | 60 | ||
| pBt087 | Forward | GACTTGTGTAACATTAGTAGGC | 62 | 598 |
| Reverse | TATACCGTATTCTTGGGATGC | 62 | ||
| pBt091 | Forward | CGATCTTAATTATGAACTCGAGC | 64 | 244 |
| Reverse | CACTTGTTTATCGTCCACTCG | 62 | ||
| pBt092 | Forward | TCAGAATTAATCAAACAAGTCGC | 62 | 257 |
| Reverse | ACAGCTTCTTTTAATGCTTTTCC | 62 | ||
| pBt093 | Forward | TCTTTGCAGGAACAATTGTTGC | 62 | 180 |
| Reverse | GAATCGAATCGTCGAGATTGC | 62 | ||
| pBt094 | Forward | GAAAGCAACCGGTATTGTACG | 62 | 270 |
| Reverse | ACTTTGTATTGTTCTAATTCTTCC | 62 | ||
| pBt097 | Forward | GGCTGAAGAGAAGGCATGGC | 64 | 343 |
| Reverse | CACAGTGCTCCTAGTCTCTCC | 66 | ||
| pBt098 | Forward | TGCCTATGAGTCATTGGCGG | 62 | 356 |
| Reverse | CTGCATATGATGCCGCTGAGC | 66 | ||
| pBt100 | Forward | CGAAAATTATATCTCCAAGATCC | 62 | 582 |
| Reverse | GCATGGAATATGCATATCACC | 60 | ||
| pBt101 | Forward | CTATATGCTAGTGCTGAAGCC | 62 | 403 |
| Reverse | CATTATCAACGATAACCAATCGC | 64 | ||
| pBt102 | Forward | AGAGGTACACGTGTTAGTACAC | 64 | 358 |
| Reverse | GTTGTGCGCCTGATGTAATCATA | 66 | ||
| pBt108 | Forward | ACGTTGTGAAACAAATTCTATGG | 62 | 303 |
| Reverse | GTTCATTCATATTACTAAGTACG | 60 | ||
| pBt120 | Forward | CTATTAGGTTCAGAAAATAATCC | 60 | 246 |
| Reverse | TAACTCTATTTCTTTGCTCATGC | 62 | ||
| pBt131 | Forward | CCATGCTTGGGATTATTATCG | 60 | 683 |
| Reverse | AGTTGCATCACTTGGCTTGCG | 64 | ||
| pBt132 | Forward | CGTATTATCAAGGAAAATTGGC | 60 | 455 |
| Reverse | CGTTGGATTATTCGCAATCGC | 62 | ||
| pBt133 | Forward | CACTAATTGTTATTGTGGCAGC | 62 | 686 |
| Reverse | TTGGATTTGTGACTTCTGAAGC | 62 | ||
| pBt136 | Forward | TGTTCGAGATTATGGGTTATTTTG | 64 | 218 |
| Reverse | CCATGCAACAGCTTGTGCTTTTA | 66 | ||
| pBt137 | Forward | CCTTATTATTTGAGGGTATTACG | 62 | 652 |
| Reverse | GCTGAAATTATGTATAAAATTAGG | 60 | ||
| pBt138 | Forward | CACATAAGTTTGAAATATATATGG | 60 | 425 |
| Reverse | GTATATTGAATTTTACGAAGCCG | 62 | ||
| pBt139 | Forward | CTACATAATATCTCATTTTCTGC | 60 | 377 |
| Reverse | GACCAATACAATTTGTGACGC | 60 | ||
| pBt145 | Forward | GAAATTAGGTGCTGGAGTGG | 60 | 446 |
| Reverse | ATTTTAATCCACCTGTTTCTGC | 60 | ||
| pBt147 | Forward | CGAAATTACAATCATTTCAAAAGG | 62 | 225 |
| Reverse | AGAATCGAATCGTAGAGATTGC | 62 | ||
| pBt148 | Forward | TGAAAGCAACAGGTATTGTACG | 62 | 252 |
| Reverse | CAGCTAACAGTTGTTTTAGACC | 62 | ||
| pBt149 | Forward | TCTATATGGATATTGAGTATTACG | 62 | 273 |
| Reverse | CCTCTATTCTCTTCATGCTCTC | 64 | ||
| pBt152 | Forward | GGAGATTTGACTGCCTTTGG | 60 | 675 |
| Reverse | CCAATCATTACAATTAACAGCG | 60 | ||
| pBt156 | Forward | AAGAGGCTGATTTATTCGCAGG | 64 | 425 |
| Reverse | GTTAATTTCATCAGGATCACCG | 62 | ||
| pBt157 | Forward | TGAATAGGGATCACTTTTATACG | 62 | 279 |
| Reverse | AACACTCTACTATTTCTACACC | 60 | ||
| pBt158 | Forward | TCTGAACTTACTGGGCTTTCG | 62 | 397 |
| Reverse | TAAGATCCTGATTTCTACGTGC | 62 |
a
Approximate denaturation temperatures (Tm) (based on approximations for short sequences) were determined as follows: Tm = 4(G + C) + 2(A + T).