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Characteristics of gene-specific primers for probe cDNA generation
| Parameter | Limits |
|---|---|
| Primer length | 16-25 nt |
| Primer Tm | 52-60°C |
| Primer dTm | ≤ 2°C |
| 3' pentamer stability | 8.5 -kc/M |
| Product size | 250-550 bp (tolerated 250-750 bp) |
| Product location | 3' proximal 1000 nt (tolerated 1500 nt) |
| Product Tm | 70-90°C |
