Hello everyone,
I work with skin cells and am performing RT-qPCR of primary fibroblast cells and primary keratinocyte cells to further use as a comparison to iPSC-derived skin cells and 3D skin in vitro model.
The main question: I would like to use TBP and RPL13A as reference genes for keratinocytes and GAPDH and RPL13A for fibroblasts. This will mean that the data is normalized to different reference genes. Is this a problem for further comparison? We are also doing normal iPSC culture characterization with RT-qPCR and not sure if then I use the combination of TBP + RPL13A or GAPDH+RPL13A. Please let me know your opinions.
Thank you!
Reference gene validation should be performed for all samples used in experiments, taking into account one reference gene. The about 6 or more reference gene panel and programs such as NormFinder are used to validate the reference gene. The program will suggest the most stable gene for all tested cells.
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