Table 2 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 |