Table 2. Primers, probes and standard curve parameters for the targets used in the conventional and real time PCR assays.
| Target | Sequence | Amplicon size | Reference | Slope | Intercept | Coefficient of determination (r2) | Amplification efficiency (%) |
|---|---|---|---|---|---|---|---|
| 18S rDNA | 18S rDNA F 5’-GTACTGGGGCGTCAGAGGT-3’ 18S rDNA R 5’-TGGGTGTCATCGTTTGCAG-3’ 18S rDNA Tq 5’- FAM AATTCTTAGACCGCACCAAG-NFQ-MGB-3' |
155 bp | Present study | -3.45 | 28.41 | 0.99 | 94.92 |
| HSP70 | HSP70 F 5'- CACCATCACCAACGACAAGG -3' HSP70 R 5'- GTCGGAGACCGTGTTCTTCA -3' HSP70 Tq 5'- FAM CTGAGCAAGGACGAGAT -NFQ-MGB-3’ |
163 bp | Present study | -3.25 | 32.52 | 0.97 | 103.09 |
| HSP70c | HSP70cF 5’-GGACGAGATCGAGCGCATGGT-3’ HSP70cR 5’-TCCTTCGACGCCTCCTGGTTG-3’ |
234 bp | Graça et al., 2012[11] | - | - | - | - |
| RNAse P–VIC/TAMRA | Not Available | Not Available | Applied Biosystems (Cat. No. 4316844) | -3.43a/-3.18b | 27.38a/27.64b | 0.99a/0.99b | 95.68%a/106.28%b |
Parameters of qPCR standard curves for both targets are showed.
aqPCR standard curve values for RNAse P in multiplex with 18S rDNA.
bqPCR standard curve values for RNAse P in multiplex with HSP70.