iTaq™ Universal Probes One-Step Kit

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iTaq Universal Probes One-Step Kit

Overview

Superior Data Precision and Reproducibility

Superior data precision and reproducibility with iTaq universal probes one-step kit

Superior data precision and reproducibility with iTaq™ universal probes one-step kit. A GAPDH assay with an input of 100–8.78 ng RNA was performed on a CFX96™ real-time PCR detection system. RFU, relative fluorescence units.

Greater Sensitivity with iTaq™ Universal Probes One-Step Kit

Duplex PGK1 assay data for iTaq

Duplex PGK1 assay data for TaqMan

Singleplex PGK1 assay data for iTaq

Singleplex PGK1 assay data for TaqMan

Greater sensitivity with iTaq universal probes one-step kit. A high-performing RT-qPCR reaction, using a PGK1 assay in duplex with a PPIA assay, with an input of 100 ng–10 pg RNA was performed on a CFX96™ real-time PCR detection system. iTaq universal probes one-step kit (A) and TaqMan RNA-to-CT 1-step kit from Life Technologies (B) were able to amplify the duplex reaction. Singleplex PGK1 assay data are shown for the iTaq (C) and TaqMan (D) kits. The iTaq kit exhibits a slight 2–quantification cycle (Cq) delay from singleplex to duplex, which is typical for duplex reactions; however, the TaqMan kit exhibits a 4-Cq delay (greater than one log of dynamic range), which indicates less sensitivity. RFU, relative fluorescence units.

Superior RT-pPCR Performance in a Triplex Reaction

Dye Logs Standard Curve, R2 Efficiency, %
 FAM 5 0.998 106
 HEX 4 0.999 101
 Quasar 6 0.99 103

 

Dye Logs Standard Curve, R2 Efficiency, %
 FAM 4 0.998 101
 HEX 4 0.993 97.6
 Quasar 4 0.998 97.5

 

Dye Logs Standard Curve, R2 Efficiency, %
 FAM 4 0.999 96.7
 HEX 4 0.999 96.7
 Quasar 4 0.999 95.6

Superior RT-qPCR performance in a triplex reaction with iTaq™ universal probes one-step kit. An XPO (FAM), TUB (HEX), and GAPDH (Quasar) assay set with an input of 100 ng–100 fg RNA was performed on a CFX96™ real-time PCR detection system. (A) iTaq universal probes one-step kit exhibited the greatest dynamic range across all three assays, which indicates a high degree of sensitivity. In addition, the iTaq kit demonstrated the lowest standard deviations for technical replicates. Brilliant III ultra-fast QRT-PCR master mix from Agilent Technologies (B) and TaqMan RNA-to-CT 1-step kit from Life Technologies (C) did not perform as well, exhibiting four logs of dynamic range for each assay (lower sensitivity) and higher standard deviations for technical replicates (lower data precision). RFU, relative fluorescence units.

Optimal RT-qPCR Efficiency and Sensitivity

iTaq universal probes one-step kit produced standard curve R2 = 0.999 and efficiency = 98.1%

Brilliant III ultra-fast QRT-PCR master mix (Agilent Technologies) produced standard curve R2 = 0.984, efficiency = 87.3%, and delayed quantification cycle (Cq) values with less than optimal data precision for the technical replicates

Optimal RT-qPCR efficiency and sensitivity with iTaq™ universal probes one-step kit. A 72% GC-rich CBP assay with an input of 100 ng–10 pg RNA was performed on a CFX96™ real-time PCR detection system. A, iTaq universal probes one-step kit produced standard curve R2 = 0.999 and efficiency = 98.1%; B, Brilliant III ultra-fast QRT-PCR master mix (Agilent Technologies) produced standard curve R2 ≥ 0.984, efficiency = 87.3%, and delayed quantification cycle (Cq) values with less than optimal data precision for the technical replicates. RFU, relative fluorescence units.

Our Universal Real-Time PCR Reagents Web App brings you in-depth product information, individual and competitive performance data, interactive tutorials, 3-D animated videos, technical manuals, and more.

The iTaq universal probes one-step kit is a fast and convenient solution for real-time PCR using the powerful combination of iScript™ RNase H+ MMLV reverse transcriptase and antibody hot-start iTaq DNA polymerase in one reaction, providing improved PCR efficiency, wider dynamic range, superior sensitivity and specificity, and inhibitor tolerance — even with cell lysates — without affecting performance.

  • Increased sensitivity, specificity, and efficiency — advanced buffering chemistry enables robust performance and increased resistance to a wide variety of sample types and target sequences
  • Obtain superior results with singleplex and multiplex reactions — advanced formulation enables the simultaneous amplification of up to two targets, resulting in higher data precision with fewer pipet steps and reduced sample usage
  • High-throughput real-time PCR screening and validation — simplified work flow and reduced cycling times enable screening and validation of the greatest number of samples and targets in the shortest period of time
  • Use any ROX-dependent or -independent real-time PCR system — the universal reference dye in this supermix enables ROX normalization of qPCR data regardless of the ROX level requirements of the qPCR system
  • Achieve superior gene expression results under various cycling conditions — our robust formulation delivers consistent performance in standard or fast cycling conditions, primer concentrations, and temperature ranges, allowing cost and time savings

Applications and Uses of the iTaq Universal Probes One-Step Kit

  • RT-qPCR/real-time PCR
  • Gene expression analysis
  • Multiplexing
  • Absolute quantification
  • Mutation detection
  • Pathogen detection
  • Viral detection (load)
  • Genetically modified organisms (GMO) characterization
  • Genetic profiling

Instrument Compatibility

The iTaq universal probes one-step kit is compatible with all real-time qPCR systems.

Related Products

We have developed iTaq™ universal SYBR® Green one-step kits available in formats of 100 reactions (171-5140) or 500 reactions (171-5141).

Bio-Rad offers a complete workflow solution for real-time PCR, including:

More Information

To ensure that your qPCR reactions are reproducible and reliable, follow our step-by-step design and optimization protocols. We also offer insights into analysis of qPCR data and troubleshooting reactions.

iTaq™ Universal Probes One-Step Kit, 100 x 20 µl reactions

172-5140
100 x 20 µl reactions, 1 ml (1 x 1 ml) 2x one-step reaction mix, containing iTaq DNA polymerase, buffers, and ROX normalization dyes; iScript™ reverse transcriptase; and nuclease-free water

List Price:   $200.00
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iTaq™ Universal Probes One-Step Kit, 500 x 20 µl reactions

172-5141
500 x 20 µl reactions, 5 ml (5 x 1 ml) 2x one-step reaction mix containing iTaq DNA polymerase, buffers, and ROX normalization dyes; iScript™ reverse transcriptase; and nuclease-free water

List Price:   $728.00
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Number Description Options
6516 Better, Faster, and Less Expensive Gene Expression Analysis Using Multiplex One-Step RT-qPCR: A Case Study of Colorectal Cancer, Rev A Click to download
6252 Reagent Comparison Guide for Real-Time PCR Brochure, Rev C Click to download
6090 Amplification Reagents and Plastics Brochure, Rev D Click to download [ Add to Cart (Free) ]
10032046 iTaq Universal Probes One-Step Kit Product Insert, Rev B Click to download