TABLE 1.
PCR primers and formulae used to determine V. cholerae VNTR repeat numbers
| Locus | Dye-primer | Expected rangea (bp) | Formulab |
|---|---|---|---|
| VC0147 | Tetc-ACGTGCAGGTTCAACCGTG | 186-224 | (x − 150)/6 |
| TTGTCATGGCTTGGATTTGG | |||
| VC0437 | Tet-GTTGCCGCCATCACCAGCTTG | 265-301 | (x − 245)/6 |
| CGTTAGCATCGAAACTGCTG | |||
| VC1650 | Tet-CCGCTAACTGAGTGACCGC | 370-440 | (x − 307)/9 |
| CTACCAAGCGGCGGTTAAGCTG | |||
| VCA0171 | Famd-AGGCGCCTGATGACGAATCC | 316-442 | (x − 270)/6 |
| GCTGAAGCCTTTCGCGATCC | |||
| VCA0283 | Fam-GGAGGTAGCTACGAATTCTAC | 118-244 | (x − 95)/6 |
| GTACATTCACAATTTGCTCACC |
a
The expected range of the sizes of the fragments produced by amplification using the primers is shown.
b
In each formula, x is the size of the fragment for each individual isolate and locus pair. Genemapper v4.0 produces sizes in hundredths of a base pair. When the formulas are applied, the value is rounded to the nearest whole number to determine the number of repeats.
c
Tet, 6-tetamidite.
d
Fam, 6-carboxyfluorescein.