Genes | Initial cycle | Denaturation | Annealing | Extension | Final extension | Primers sequencing | Product size (bp) | |
---|---|---|---|---|---|---|---|---|
sfa gene | ||||||||
Time | 4 M | 30 S | 30 M | 40 S | 3 M | F:5′ C C G T A A G A T G T C T G C G A G 3′ | R:5′ A G C A A G T C T G G C A A C G A G 3′ | 100 |
Temperature | 95 | 95 | 53 | 72 | 72 | |||
Number of cycles | 1 | 35 | 35 | 35 | 1 | |||
afa gene | ||||||||
Time | 5 M | 1 S | 30 M | 3 S | 7 M | F:5′ G C T G G G C A G C A A A C T G A T A A C T C T C 3′ | R:5′ C A T C A A G C T G T T T G T T C G T C C G C C G 3′ | 750 |
Temperature | 95 | 95 | 60 | 72 | 72 | |||
Number of cycles | 1 | 35 | 35 | 35 | 1 | |||
foc gene | ||||||||
Time | 4 M | 1 S | 1 M | 2 S | 10 M | F:5′ G G T G G A A C C G C A G A A A A T A C 3′ | R:5′ G A A C T G T T G G G G A A A G A G T G 3′ | 388 |
Temperature | 95 | 95 | 58 | 72 | 72 | |||
Number of cycles | 1 | 35 | 35 | 35 | 1 |