In molecular biology, the polymerase chain reaction or (PCR) is a technology, which is used to intensify a single copy or a few copies of a piece of DNA across various orders of magnitude, producing millions of copies of a certain DNA sequence. Read this article further to learn what are the 4 risk factors or compliance issues in PCR assays.
1. Handling Reagents
Right handling of reagents is an important factor in PCR. Here are a few tips to handle reagents: Track everything that makes up the mix, which includes even water. One must quickly be able to identify any and all components of a PCR master mix. This permits one to determine simple technician error from reagent issues. Without this, you won’t be able to track issues in reagents.
2. How to Track Reagents
Log everything that comes into the laboratory itself, not what’s received. Aliquot reagents as quickly as possible. Keep reagents in as small a quantity as feasible. It’s less risky to grab multiple tubes of reagents than to dip in reagent tubes for different set-ups. Test new aliquots promptly in a new, well-qualified reaction. Goal is to compare old reagents to new.
3. Tracking Water in the Mix
Water can easily be contaminated with microbes that can potentially create false signals. People tend to be more cavalier with handling water as compared to other reagents. It is also the easiest reagent to get rid of, so starting there always helps.
4. Handling Consumables
Never, ever, store consumables in a reaction set-up area because these areas are sources of contamination and PCR inhibition issues. Try to make sure each technician has his/her own set of supplies so that it would be easier to recognize technician error this way. If possible, keep consumables for different reactions separate to avoid cross-contamination.