Why am I seeing product in my no template control (NTC)?
The most likely cause of product in the negative controls is contamination. Here are a few suggestions to prevent contamination: We highly recommend that you aliquot your probe and primers into experiment size volumes. This will decrease the chances of contamination, allow you to start with a fresh tube if you do have contamination, and will minimize freeze thaw cycles that can degrade oligonucleotide quality. Use dedicated work areas for setting up the PCR set-up, adding the DNA/template, and handling the amplification product. Clean PCR work areas regularly with 70% ethanol and UV irradiation. Pipettes: Use 70% ethanol to clean micropipettes regularly. Use sterile and filtered pipette tips to minimize aerosols. We recommend having separate pipettes and pipette tips for PCR set-up and post-PCR analysis. If you do find contamination in your sample be sure to replace all reagents and stock buffers and thoroughly clean the PCR preparative areas.