Hello,
I am just curious about the comparability of results obtained from Taqman vs SyBR Green chemistry.
i.e. If I do one experiment using Taqman and another with SyBR, would those results be comparable or can only experiments done using the same method be compared to each other.
Thanks,
Alex
In general, these two methods are comparable if your primers are designed properly.
Dye-based assays are perfect for many real-time qPCR applications. When you understand the true advantages and disadvantages of probe-based assays, you can reduce the cost while keeping the quality of your qPCR experiments high.
Novice qPCR users often argue that data generated with dye-based assays are susceptible to misinterpretation: the detection of nonspecific PCR products would influence the Cq values.
But the detection of nonspecific PCR products should not be cancelled out by adding a probe to the qPCR reaction. It is through careful experiment design and optimization that you obtain specific, efficient and sensitive amplification and detection of your target of interest. Primer concentrations, denaturation and annealing times and temperatures, and buffer composition are few of many critical parameters that contribute to a robust qPCR reaction. When sensitivity is not an aspect that you want to improve, optimizing these parameters will give you a suitable qPCR assay. And it does not need a probe. This is the strategy applied by Biogazelle when designing Bio-Rad's PrimePCR assays: the most comprehensive portfolio of validated Research Use Only dye-based qPCR assays for human, mouse and rat.
You can find additional arguments, and additional advice on when to use what detection technology at https://blog.qbaseplus.com/taqman-probe-or-sybr-green-dye
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