The relative standard curve method in qPCR is a standard curve-based method for relative real time PCR data processing. Can anyone explain the calculations of this method to me? I read something about pooling of cDNA samples and using them as a calibrator is this right?
Dear Omnia Adel Badr, this topic is quite wide. The best storage of appropriate data is on https://www.gene-quantification.de/ reading these articles and guides is very useful. But I understand that it is very laborous. In two words...
1. Standard curve means serial dilutions of your target genes. One of the possible ways is cDAN/DNA (depends on your start material and goals) serial dillution. For example 5-fold (you can also use 2-fold or 10-fold, but for me 5-fold is better). It is callibration curve that can provide you quantitative information about target gene copy number in your samples (depends on your choise: cells, genome equivalents, ng/ul concentration and oth).
2. Relative means any relative quantity. For example (it is very popular) your target gene copy number (on the base on standard curve) in comparison to some reference genes that don't affected under experimental conditions (it is not so easy to detemine it )).
3. Relative standard curve means at least two curves: for target gene and for reference gene. After that you can compare quantity and receive relative quantity. For exapmle 10000 copies or target gene and 100000 copies of reference gene in your control sample (native, non cancerous tissue for ex.) and for your sample under inve
Dear Andrei
Many thanks for your clear and valuable answer. Your explanation for the concept of Relative standard curve is fantastic and I will read the guide articles for more understanding. I appreciate your help so much.
Regards,
Omnia
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