Table 2.
DNA regions targeted for PCR amplification in this study; original names, sequences, and literature sources of the oligonucleotide primers used; and values for the amplification reaction conditions that varied among primer sets
| Targeta | Primer name | Sequence (5′→3′) | Reference | Primer concn (μM) | MgCl2 concn (μM) | Annealing temp (oC) |
|---|---|---|---|---|---|---|
| Bacterial (rrn) 16S rRNA | 27B Fb | AGRGTTYGATYMTGGCTCAG | 47 | 0.6 | 15 | 50 |
| 1492 R | GGYTACCTTGTTACGACTT | 38 | ||||
| Vc (ITS) intergenic transcribed spacer | prVC F | TTAAGCSTTTTCRCTGAGAATG | 10 | 0.6 | 15 | 60 |
| prVCM R | AGTCACTTAACCATACAACCCG | |||||
| Vc (ctx) cytotoxin | P1 | TGAAATAAAGCAGTCAGGTG | 36 | 0.5 | 15 | 55 |
| P2 | GGTATTCTGCACACAAATCAG | |||||
| Vp (tlh) thermolabile hemolyisn | L-tl | AAAGCGGATTATGCAGAAGCACTG | 5 | 0.5 | 25 | 58 |
| R-tl | GCTACTTTCTAGCATTTTCTCTGC | |||||
| Vp (tdh) thermostable direct hemolysin | TDH F | GTAAAGGTCTCTGACTTTTGGAC | 5 | 0.5 | 25 | 55 |
| TDH R | TGGAATAGAACCTTCATCTTCACC | |||||
| Vp (trh) tdh-related hemolysin | TRH F | TGGGCTTCGATATTTTCAGTATCT | 5 | 0.5 | 25 | 55 |
| TRH R | CATAACAAACATATGCCCATTTCCG | |||||
| Vv (vvhA) hemolysin | L-CTH | TTCCAACTTCAAACCGAACTATGAC | 7 | 0.6 | 15 | 63 |
| R-VVH | ATTCCAGTCGATGCGAATACGTTG | 55 | ||||
| Vv (rrn) 16S rRNA | Vib 1 | GTGGTAGTGTTAATAGCACT | 34 | 1 | 15 | 55 |
| Vib 2 | TCTAGCGGAGACGCTGGA | |||||
| Vib 3R | GCTCACTTTCGCAAGTTGGCC |
a
Presumed specificity is indicated by the abbreviations Vc (V. cholerae), Vp (V. parahaemolyticus), and Vv (V. vulnificus), followed by the gene (or region) abbreviation and a brief description of the function of the target.
b
Referred to in the cited paper as a variation of 16S rRNA bacterial primer 8F. The new name reflects the convention of designating the primer by the position of the 3′ rather than the 5′ end, and the “B” indicates a change in the degeneracy compared to the conventional 16S rRNA 27F primer.