Table 2. Comparison of parameters of four tested RT-qPCR methods for miRNA quantification. Cost estimate per assay with 20 RT and 60 qPCR reactions is indicated (details available in suppl. file).
| cDNA synthesis strategy | Sensitivity (miRNA copies in analyzed volume) | Linear dynamic range | Unspecific cross-reaction | Accurate detection of isomiRs | Melting curve for specificity control | Detection of piRNAs and plant miRNAs | Cost | |
|---|---|---|---|---|---|---|---|---|
| Two-tailed RT-qPCR | specific RT primer | 102 - 103 | 6–7 logs | < 1% | yes | yes | yes |
|
| TaqMan | specific RT primer | 102 - 103 | 6–7 logs | < 23% | no | no | yes |
|
| Quanta | poly(A) tailing | 102 - 103 | 6–7 logs | < 51% | yes | yes | no |
|
| miQPCR | linker ligation | 104 | 5 logs | > 100% | no | yes | yes, but efficiency is reduced |
|