iTaq™ Universal SYBR® Green One-Step Kit

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iTaq Universal SYBR Green One-Step Kit

Overview

Superior 1.5-fold Precision
The iTaq™ universal SYBR<sup>®</supb> Green one-step kit demonstrated robust data acquisition even with small-fold changes.

 

The Qiagen QuantiTect SYBR<sup>®</supb> Green RT-PCR kit and Life Technologies Power SYBR<sup>®</supb> precision RNA-to-Ct kit

 

Both showed less than optimal precision. ΔRn, baseline-corrected normalized reporter.

Superior 1.5-fold precision with iTaq™ universal SYBR® Green one-step kit. A human TUBA assay with an input of 100 to 8.78 ng total RNA was performed on an ABI 7900 real-time instrument. A, The iTaq™ universal SYBR® Green one-step kit demonstrated robust data acquisition even with small fold changes. B, The Qiagen QuantiTect SYBR® Green RT-PCR kit and Life Technologies' Power SYBR® precision RNA-to-Ct kit (C) both showed less than optimal precision. ΔRn, baseline-corrected normalized reporter.


Superior Dynamic Range and Sensitivity Performance

 


 


Superior dynamic range and sensitivity performance of iTaq™ universal SYBR® Green one-step kit. A cell lysate and 226 bp human GAPDH assay were used to determine dynamic range and sensitivity performance on a CFX384™ real-time PCR instrument. A, The iTaq™ universal SYBR® Green one-step kit demonstrated 1,000 to 0.001 cell input, a PCR efficiency of 96.8%, and R2 of 0.997. Both competitor mixes showed less than ideal performance; B, the Qiagen QuantiTect SYBR® Green RT-PCR kit amplified from 1,000 to 1 cell input range with a PCR efficiency 94.5% and R2 of 0.977; C, the Power SYBR® RNA-to-Ct kit amplified from 100 to 0.1 cell input range with a PCR efficiency of 83.3% and R2 of 0.981. RFU, relative fluorescence units.


Superior RT-qPCR Inhibitor Tolerance

 


 


Superior RT-qPCR inhibitor tolerance of iTaq™ universal SYBR® Green one-step kit (). A human ACTB assay was performed with 200 ng of total RNA, and chocolate and heparin were spiked in to evaluate the tolerance. A, Chocolate spiked in from 0.01 to 0.2% resulted in a greater quantification cycle (Cq) delay from the control sample with the Life Technologies Power SYBR® RNA-to-Ct kit indicating less tolerance for RT-qPCR inhibitors; B, heparin spiked in from 200 to 3,200 pg/µl caused the 17.83 Cq delay at 200 pg/µl () and eventual complete inhibition with the Life Technologies Power SYBR® RNA-to-Ct kit, but only minor Cq delays with the iTaq™ universal SYBR® Green one-step kit; C, heparin spiked in from 200 to 1,600 pg/µl produced a greater Cq delay from the control sample with the Qiagen QuantiTect SYBR® Green RT-PCR kit ().


GC-Rich Template

 


Superior dynamic range and sensitivity performance of iTaq™ universal SYBR® Green one-step kit. A 93-bp human BAX assay was performed with 500 ng to 5 pg of total RNA on an AB ViiA 7 real-time PCR instrument. A, The iTaq™ Universal SYBR® Green one-step kit gave a PCR efficiency of 102.4% and R2 of 0.999; B, the Life Technologies Power SYBR® RNA-to-Ct kit yielded a PCR efficiency of 111.8% and R2 of 0.992. The delayed cycle threshold (Ct) values, loss of dynamic range and the 500 ng input, and compression of the 50 ng input in the Power SYBR® RNA-to-Ct kit demonstrated RT-qPCR inhibition at higher RNA inputs and lack of low-level target expression detection. ΔRn, baseline-corrected normalized reporter.


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 SYBR® Green 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 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.

  • Enhanced efficiency, specificity, and sensitivity — our patented* inhibitor reducer prevents the interference of the RT enzyme, which leads to reduced sensitivity and efficiency, with the DNA polymerase during the replication of the cDNA
  • Use any ROX dependent or independent real-time PCR systems — 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, sample and target types, and temperature ranges, allowing cost and time savings
  • Obtain better results with qPCR assays — simplify your workflow and experimental design using our laboratory-validated PrimePCR™ assays (currently available in the U.S., Canada, and E.U. only)

* U.S. patent 8,338,094.

Applications and Uses of the iTaq™ Universal SYBR® Green One-Step Kit

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

Instrument Compatibility

The iTaq™ universal SYBR® Green one-step kit is compatible with all Bio-Rad real-time qPCR systems.

Related Products

We have developed iTaq™ universal probe one-step kits, available in formats of 100 reactions (172-5140) or 500 reactions (172-5141).

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.

Product expiration (after shipping date) 12 months
RT storage at –20°C (after shipping date) 12 months
Reaction mix storage at 4°C (after thawing) Up to 3 months
Customer guarantee (minimum usage time) at –20°C 12 months
iTaq™ Universal SYBR® Green One-Step Kit

172-5150
100 x 20 µl reactions, 1 ml (1 x 1 ml), one-step RT-PCR kit, includes 2x one-step SYBR® Green reaction mix containing iTaq DNA polymerase, buffers, and ROX normalization dyes; iScript™ reverse transcriptase; and nuclease-free water

List Price:   $189.00
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iTaq™ Universal SYBR® Green One-Step Kit

172-5151
500 x 20 µl reactions, 5 ml (5 x 1 ml), one-step RT-PCR kit, includes 2x one-step SYBR® Green reaction mix containing iTaq DNA polymerase, buffers, and ROX normalization dyes; iScript™ reverse transcriptase; and nuclease-free water

List Price:   $691.00
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Number Description Options
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) ]
10032048 iTaq™ Universal SYBR® Green One-Step Kit Product Insert, Rev B Click to download
6290 PrimePCR Assays and Panels Brochure, Rev C Click to download