Table 1.
Modified Tissue Processing Protocol for Preservation of RNA in Ethanol-Fixed Tissues
| Tissue processing step | Duration, minutes | Temperature, °C |
|---|---|---|
| 70% Ethanol | 10 | 35† |
| 80% Ethanol | 10 | 35† |
| 95% Ethanol* | 15 | 35† |
| 100% Ethanol* | 15 | 35† |
| Xylene | 10 | 35† |
| Xylene | 15 | 35† |
| Low-melt paraffin | 15 | 56‡ |
| Low-melt paraffin | 15 | 56‡ |
| Low-melt paraffin | 15 | 56‡ |
| Low-melt paraffin | 15 | 56‡ |
| Total minutes | 135 |
*Single 95% and 100% ethanol steps rather than conventional duplicate steps for each.
†Temperature reduced from 40°C to 35°C.
‡Low-melting temperature paraffin steps at 56°C for 60 minutes instead of standard 58°C incubations for 120 minutes. Processing time reduced from the conventional 450 total minutes to 135 minutes.