In the reverse transcription (RT) step, cDNA is reverse transcribed from total RNA samples using a small RNA-specific, stem-loop RT primer from the TaqMan® Small RNA Assays and reagents from the TaqMan® MicroRNA Reverse Transcription Kit.
Prepare the qPCR reactions in an area free of artificial templates.
When preparing samples for PCR amplification:
- Use a positive-displacement pipette or aerosol-resistant pipette tips.
- Follow proper pipette-dispensing techniques to prevent aerosols.
- Wear clean gloves and a clean lab coat (not previously worn while handling amplified PCR products or used during sample preparation).
- Change gloves whenever you suspect that they are contaminated.
- Maintain separate areas and dedicated equipment and supplies for:
- Analysis of PCR products
- Never bring amplified PCR products into the PCR setup area.
- Open and close all sample tubes carefully. Centrifuge tubes before opening. Try not to splash or spray PCR samples.
- Keep reactions and components capped as much as possible.
- Clean lab benches and equipment periodically with 10% bleach solution.