Table 2. Primers and conditions for Trypanosoma cruzi molecular typing.
Target | Primers | Sequence | Concentration | Termocycling | |||
---|---|---|---|---|---|---|---|
D | A | E | C | ||||
SL-IR I and II | TCC TC2 TC1 |
CCCCCCTCCCAGGCCACACTG CCTGCAGGCACACGTGTGTG TCCGCCACCTCCTTCGGGCC |
2 µM 2 µM 2 µM |
94°C/1’ 94°C/30’’ 94°C/1’ 94°C/1’ |
67°C/1’ 65°C/1’ 63°C/1’ 61°C/1’ |
72°C/1’ 72°C/1’ 72°C/1’ 72°C/1’ |
5 5 5 30 |
24Sa-rDNA First round Second round |
D76 D75 D76 D71 |
GGTTCTCTGTTGCCCCTTTT GCAGATCTTGGTTGGCGTAG GGTTCTCTGTTGCCCCTTTT AAGGTGCGTCGACAGTGTGG |
0,5 µM 0,5 µM 0,5 µM 0,5 µM |
94°C/1’ 94°C/1’ 94°C/1’ 94°C/1’ |
60°C/1’ 57°C/1’ 55°C/1’ 58°C/1’ |
72°C/1’ 72°C/1’ 72°C/1’ 72°C/1’ |
5 5 5 35 |
A10 First round Second round |
Pr1 P6 Pr1 Pr3 |
CCGCTAAGCAGTTCTGTCCATA GTGATCGCAGGAAACGTGA CCGCTAAGCAGTTCTGTCCATA CGTGGCATGGGGTAATAAAGCA |
0,5 µM 0,5 µM 0,5 µM 0,5 µM |
94°C/1’ 94°C/1’ 94°C/1’ 94°C/1’ 94°C/1’ |
60°C/1’ 57°C/1’ 55°C/1’ 58°C/1’ 65°C/1’ |
72°C/1’ 72°C/1’ 72°C/1’ 72°C/1’ 72°C/1’ |
5 5 5 35 35 |
D: denaturation, A: annealing, E: extension, C: cycles.