Suggested Technical Recommendations for Improved RT-PCR of PML/RAR
| Steps . | Methods . | Advantages . | Reference . |
|---|---|---|---|
| RNA extraction | Rapid suspension of MNC in GTC (diagnostic samples) | Less frequent RNA degradation | 19 |
| Ethidium bromide gel visualization | Identification of possible false-negative | 97 | |
| RT | Increased RNA amount | ||
| RNA denaturation Prolonged incubation | Increased cDNA yield | 89 | |
| Amplification | Hot-start PCR (or Gold Taq polymerase) | One-round (not nested) PCR for diagnosis; increased specificity (cleaner products); increased sensitivity | 90 |
| Steps . | Methods . | Advantages . | Reference . |
|---|---|---|---|
| RNA extraction | Rapid suspension of MNC in GTC (diagnostic samples) | Less frequent RNA degradation | 19 |
| Ethidium bromide gel visualization | Identification of possible false-negative | 97 | |
| RT | Increased RNA amount | ||
| RNA denaturation Prolonged incubation | Increased cDNA yield | 89 | |
| Amplification | Hot-start PCR (or Gold Taq polymerase) | One-round (not nested) PCR for diagnosis; increased specificity (cleaner products); increased sensitivity | 90 |
Abbreviations: MNC, mononuclear cells; GTC, guanidium isotiocyanate.