TABLE 3.
Primer sequences and CFLP conditions for doubly fluorescence-labeled PCR products from target genes adk, aspC, carA, dhp, and glnA
| Gene | Fluorescence-labeled primers (sequences)a | PCR annealing temp (°C) | PCR product (bp) | CFLP digestion conditions (°C/min) |
|---|---|---|---|---|
| adk | TET (AC TTC GGT TTG CTC GTG GTA) | 68 | 479 | 58/2 |
| FAM (GG CAA AGG CAC TCA GGC G) | ||||
| aspC | TET (AA CCC WAC CGG TAT CGA CCC TAC GC) | 58 | 470 | 55/2 |
| FAM (GT CAA GCC GCT GAA AGA GAA CAT GCC) | ||||
| carA | TET (GA GTG GAT GCC CGC CTG TTC) | 65 | 919 | 62/4 |
| FAM (AT TTG GAT TAC GCC GCC CTA C) | ||||
| dhp | TET (TG CTA CTT CAC CGA CAA CAT) | 65 | 400 | 65/4 |
| FAM (TC TCA CGC CCG ATT CTT TKT) | ||||
| glnA | TET (CG TAC AGG TTT TTA TCG GCA GG) | 58 | 1,104 | 60/1 |
| FAM (AA GGC AAG CAG CAC CAC TTT AC) |
a
W, equal mixture of A and T; K, equal mixture of G and T.