Greeting everyone,
I want to know "Is there any difference in the primers of conventional PCR and qPCR?".
Thank you.
The primers are the same, but you need to be careful in how you design them. You need reactions to work very efficiently (no stable primer hairpins, no stable primer-dimers) and specifically. It will also depend on your chemistry. If you are using fluorescent dyes (SYBR, EvaGreen etc) then you need to ensure you don't get primer-dimers, because they will bind the dye and give inaccurate readings. I recommend "11 golden rules for Quantitative RT-PCR" Udvardi et al, 2008, Plant Cell as a useful reference. The primer tools on the IDT website are great for evaluating your primers.
Good luck
Hello! Yes, there can be differences between the primers used in conventional PCR and qPCR.
In conventional PCR, the goal is to amplify DNA fragments, and the primers used are designed to specifically bind to the ends of the target DNA sequence being amplified. The primers used in conventional PCR can vary in length and sequence depending on the specific target sequence being amplified and the PCR conditions being used.
In contrast, qPCR (also known as real-time PCR) is a quantitative technique used to measure the amount of DNA in a sample. The primers used in qPCR are designed to specifically amplify a target DNA sequence, but they also include additional sequences that allow for the detection and quantification of the PCR product during amplification. These additional sequences can include fluorescent dyes or probes that bind specifically to the PCR product and produce a signal that can be measured in real time.
Overall, the primers used in qPCR are designed with additional considerations for detection and quantification that are not necessarily present in conventional PCR primers.
These video playlists might be helpful to you:
https://www.youtube.com/playlist?list=PLzBiAPKi8vONSqjcweFWHMRy13y3484bE
https://www.youtube.com/playlist?list=PLzBiAPKi8vOMheKwb0mu1dcZRaeGUFSlw
https://www.youtube.com/playlist?list=PLzBiAPKi8vOPbE7Db9mKwloe34E8rXXEU
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