I recently purchased MCE's siRNA kit and encountered two questions in the experimental design. I would like to ask you: The experiment plans to use a negative control siRNA labeled with FAM. In this case, is it necessary to set up an additional negative control without a fluorescent label?
In addition to detecting its expression level by WB and QPCR, can the GAPDH positive control in the kit be detected by flow cytometry?
This may be a very simple question, but any suggestions would be greatly appreciated.
Our (MedChemExpress, MCE) siRNA kits always claim to "Three Guarantees One".
It means to contain two negative control siRNAs, one with a FAM label and the other without a FAM label. Among them, the negative control with a FAM label can be observed by fluorescence microscopy to see whether it has been successfully transfected into the cells. From the perspective of experimental design, it is recommended to set up both negative controls.
Regarding the detection of GAPDH positive control, we usually recommend the use of qPCR method. This is because our siRNA mainly acts on mRNA; while the detection of protein levels has cycle differences, and the transcripts corresponding to different targets are different, and their effects on proteins may also be different.
The answer to this question comes from MedChemExpress (MCE) Technical Support.
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