Dear Haider,
In short, yes.
Long answear: What makes a study quantitative vs. semi quantitative depends on if you know what the input amount of template is.
For example let's say you have a sample of macroalgae, dislodge all the cells on it's surface, extract community DNA and then run a PCR with the same primer set targeting a particular species, your results are:
- presence/absence if you have no idea how many cells/weight or surface area of macroalgae you started with.
- semi-quantitative if you have a rough idea of the amount of sampling material (eg. Approximately 200g macroalgae).
- quantitative if you weighted the macroalgae pr have a total cell count for every sampling point.
Yes the same primer can be used for both semi-quantitative and quantitative (real-time) PCR. There is no specific size which can be used for quantitative PCR but try to design a primer on an around 100 bp. As far as our experience the primer with amplicon length of this size works well in real-time and same can be used for normal PCR as well. Before proceeding to the real-time PCR with the primer which worked well in conventional PCR, its better to determine PCR efficiency with a standard dilution.
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