General recommendations for quality assurance in PCR analysis
Need of 3 independent areas or rooms: Area 1: Handling of reagents and preparation of amplification mixture ("Clean-room") Area 2: Sample work-up and preparation of nucleic acid, setting up PCR reactions ("Pre-amplification area") Area 3: Amplification and detection of nucleic acids ("Post-PCR analysis")
Unidirectional workflow from area 1 and 2 to 3 ("clean" work and pre-amplification first).
All rooms should have assigned equipment and reagents for the respective areas (e.g. refrigerator, vortex mixer, centrifuges, tube racks, pipette, filter tips holders etc.).
Reduction of the risk of contamination, decontamination of working areas; use of gloves, plugged pipette tips etc.
Use of DNase-free materials.
Set up the PCR under a fume hood or laminar air flow box.