Can I use conventional PCR primers for qPCR for virus quantification? i don t have any experience for qPCR and I got good band by using those primers in conventional PCR , MCP gene was cloned into plasmids and plasmid DNAs was extracted.i wanna to know is it possible I use conventional PCR primers for qPCR ?I am waiting for your opinion and comments.
Hi, Maryam, theoretically yes, you can use. But it can dependent form your primers details: length of the fragment, temperature etc.. The main point is the fragment length; for qPCR fragment size must be quite short; 100.150 bp.
You can also look on previous discussion on the same topic: https://www.researchgate.net/post/Can_primers_for_RT-PCR_be_used_to_do_the_SYBR_fluorescence_quantitative_qPCR
Hi, Maryam, theoretically yes, you can use. But it can dependent form your primers details: length of the fragment, temperature etc.. The main point is the fragment length; for qPCR fragment size must be quite short; 100.150 bp.
You can also look on previous discussion on the same topic: https://www.researchgate.net/post/Can_primers_for_RT-PCR_be_used_to_do_the_SYBR_fluorescence_quantitative_qPCR
I would say it is indeed possible, if the fragment is not too long, as stated above. You can adapt your elongation step of the qPCR accordingly. However, optimalization of your qpcr conditions is recommended, which can be done by testing a dilution series, to obtain around 100% efficiency. Despite usage of conventional primers being possible, designing specific qPCR primers might be the most straighforward way to tackle this.
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