My question is about PCR. Simple, nothing special, genotyping PCR. When I’m performing the reaction on my bench, the NTC appears contaminated. On the bench in another lab, with the same pippetors and reagents – everything is clean. Does anyone have an idea how to clean air, around a bench?
UV the entire room would probably solve your problem. Since that solution is not feasible, you can try to clean everything with 10% domestic bleach (0.5% sodium hypochlorite) including pipettes, racks, benches, centrifuges, etc., and also get new lab coat. However, if you have the chance, to avoid future contaminations you should consider the use of different laminar flow hoods for mix preparation and DNA addition, and also different rooms for pre-PCR, PCR and post- PCR. Good luck!
UV the entire room would probably solve your problem. Since that solution is not feasible, you can try to clean everything with 10% domestic bleach (0.5% sodium hypochlorite) including pipettes, racks, benches, centrifuges, etc., and also get new lab coat. However, if you have the chance, to avoid future contaminations you should consider the use of different laminar flow hoods for mix preparation and DNA addition, and also different rooms for pre-PCR, PCR and post- PCR. Good luck!
You might want to work in the laminar flowhood if there is one in the lab, else i agree with zemin wang and Conceição. Also try to change your set of buffers, MgCl2, and the other reagents if you get a band in your NTC. Hope this helps!
Several hints. First, check your reagents. Second, check pipettes, never usу the same ones for PCR and for isolation procedures. Third, wipe all surfaces on your bench with sodium hypochlorite solution , 5 min enough. Last one you may use Uracyl DNA glycosylase for PCR. Good luck.
You must routinely to check filter (Hepa Filter) or airflow in Bench, if your bench have expected of contaminant.. I hope can help you...
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