The Applied Biosystems® TaqMan® RNA-to-Ct™ 1-Step Kit delivers consistent RNA target quantification for a wide variety of assays and is validated with Applied Biosystems® comprehensive library of TaqMan® Gene Expression Assays. The TaqMan® RNA-to-Ct™ 1-Step Kit is recommended for a variety of applications including general gene expression studies, biomarker discovery, and microarray validation.

  • Accurate across a wide dynamic range for dependable performance
  • High sensitivity, enabling low copy number detection and precise quantification
  • Validated with TaqMan® Gene Expression Assays for easy experimental setup
  • Consistent performance with a wide variety of targets, including AT-rich, GC-rich and long sequences
  • Fewer pipetting steps means less hands-on-time and less chance for errors

Efficiency across a Broad Range of Input Concentrations

Maximum flexibility in RNA template input quantity requires optimal real-time PCR efficiency across a broad range of template quantities.

 Figure 1. The TaqMan® RNA-to-Ct™ 1-Step Kit (blue) shows a wider dynamic range relative to the TaqMan® One-Step RT-PCR Master Mix Reagents (orange) and comparable performance to the TaqMan® RNA-to-Ct™ 2-Step Kit (red). Amplifications are from a serial dilution of PPIA target using 100 ng to 0.1 pg of RNA input on the Applied Biosystems® 7900HT Fast Real-Time PCR System.

High Sensitivity at Low Target Concentration

The sensitivity of TaqMan® RNA-to-Ct™ 1-Step Kit was validated using a synthetic RNA template for which copy number is precisely known. Significant sampling error occurs when measuring low quantities of target, so statistical analysis is required for proper evaluation using multiple replicates. Figure 2 shows the expected quantity of target and corresponding mean Ct values. Statistical analysis indicates high confidence of sample quantification based on a T-test (Table I), consistent with as few as 10 copies of target.

Table 1: High Confidence Sample Quantification

Nominal Copies Mean Ct t-value p-value Confidence
10
20
35.48
34.55
3.34
3.34
< 0.00199.9%
20
30
34.55
33.88
3.34< 0.00199.9%
30
40
33.88
33.51
3.34< 0.00199.9%
40
50
33.51
33.15
3.34< 0.00199.9%
50
100
33.15
32
3.34< 0.00199.9%

High Sensitivity - Even With as Few as 10 - 100 Copies

Figure 2. Mean Ct obtained from amplification of 10 to 100 copies of RNA Control 250 in 32 replicates using the TaqMan® RNA-to-Ct™ 1-Step Kit on the Applied Biosystems® 7900HT Fast Real-Time PCR System.