Human TBP (TATA-box binding Protein) Endogenous Control (VIC™/TAMRA™ probe, primer limited) - FAQs

3 个常见问题解答

Can TaqMan Gene Expression Assays be used in a multiplex reaction?

Yes. For duplexing, we have VIC-labeled probes with primer limited concentrations (VIC PL) available from our pre-designed assay selection. The primer limited assay is used to detect the most abundant gene. For guidance on multiplexing, see the TaqMan Multiplex PCR Optimization User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/taqman_optimization_man.pdf).

How can I calculate the concentration of a TaqMan probe?

The initial concentration or total yield of your synthesized probe is listed on the data sheet supplied with the probe. TaqMan probes are sometimes shipped in the lyophilized state, but more often are shipped in solution (1X TE). To create a working stock solution of your probe or primers, you should reconstitute or dilute the probe in the appropriate volume of sterile 1X TE (1 mM Tris, 0.1 mM EDTA, pH 8.0) or sterile, nuclease-free H2O. Detailed instructions on how to calculate the proper volume of solution to add to your probe are provided in our online Tutorial document "Reconstituting and Diluting Primers and TaqMan Probes". You can find a copy on our website by entering this title in the main Search field.

Why does a TaqMan TAMRA probe need to have more C's (Cytosines) than G's (Guanidines) in the sequence?

Empirical data has shown that TaqMan TAMRA probes which contain more Gs than Cs will generally have lower ΔRn values and overall poor performance compared to those that do not. For more information on designing TaqMan TAMRA probes, please consult tutorials related to the Primer Express software.