TaqMan Assays を選択する理由

TaqMan probes provide better sensitivity and precision. Comparison of two 5´ nuclease PCR assays for 18S rRNA.Ten-fold dilutions of Universal Human Reference RNA (10–10^–5 ng) were prepared and analyzed in 11 replicate real-time PCR reactions using either the TaqMan Gene Expression Assay (FAM dye–labeled, with NFQ) or the non-MGB assay (FAM dye–labeled, with BHQ).Real-time PCR was run according to the respective manufacturers’ recommended conditions.Across a 6-log range of input template, the TaqMan Assay displayed earlier C_t values and better reproducibility across all data points.In addition, the TaqMan Assay had higher signal and lower background, resulting in better sensitivity and higher precision. 

Sensitivity and wide dynamic range. Sequential 10-fold dilutions of synthetic sense RNA corresponding to 4 genes—CDC23, DDX39, DCDC2, and BIRC5—were added to a background of yeast RNA to evaluate the sensitivity and dynamic range of TaqMan Gene Expression Assays.Samples containing 50 to 5 x 10⁹ target molecules were reverse transcribed, and 20% of each RT reaction was used in quadruplicate PCR reactions using Applied Biosystems™ TaqMan® Gene Expression Master Mix.Reactions containing 10 copies were detected (C_t ~35).

Reliable performance and wide dynamic range. TaqMan Gene Expression Assays were used to analyze expression of 60 targets across a 2-fold dilution series of universal reference cDNA, from 4 ng/μL to 4 pg/μL.The average slope of the lines is 1.02.TaqMan Assays exhibit virtually 100% amplification efficiency at each cycle of PCR: each target molecule is copied, doubling the fluorescence signal.