While working with the AmpliSens® GM Plant-1-FL and AmpliSens® DNA-sorb-C-M kits, I encountered the following technical issue. I performed PCR analyses to detect P-35S, T-NOS, P-FMV, and plant-specific genes. As a result, amplification for the plant genes (in the HEX channel) was observed at cycles 35, 36, and 37.
In this regard, I would like to ask the following question: what steps can be taken to achieve amplification at earlier cycles (e.g., at or before cycle 30)? Could you provide scientific recommendations regarding improvements to the DNA extraction technique, sample preparation process, or PCR conditions?
Dear Sobirjon, PCR optimization is a routine experiment and can be done using Taguchi's design. Template DNA, primers, probes, Taq poly, Mg2+, and dNTPs concentrations are subject of such optimization, as well as temperature and time of PCR cycles.
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