One-step reverse transcription (RT)-PCR kit developed specifically for SYBR® Green-based real-time PCR. Includes enzymes and reagents that reduce nonspecific amplification, providing high sensitivity, reliability, and data accuracy for qPCR.

Product Description

The Power SYBR® Green RNA-to-CT™ 1-Step Kit provides high sensitivity and specificity in an easy-to-use one-step reverse transcription-PCR kit. A novel formulation helps reduce false-positive results caused by primer-dimers that are a risk in SYBR® Green-based real-time PCR. This improves data accuracy and enables the reliable detection of low expressing targets or targets present at low amounts

  • Minimized hands-on time with a one-step protocol
  • Reduced error and contamination caused by multiple pipetting steps
  • Accuracy across a wide dynamic range of input target quantities for dependable performance
  • Highly specific and consistent results, even at low quantities of target RNA

Formulation

With the Power SYBR® Green RNA-to-CT™ 1-Step Kit you can perform reverse transcription and PCR all in one reaction tube. It includes:

  • AmpliTaq Gold® DNA Polymerase UP (Ultra Pure) for hot-start PCR to prevent amplification during reaction setup
  • ArrayScript™ UP Reverse Transcriptase for highly-efficient reverse transcription across a wide range of targets
  • A dNTP blend including dUTP which makes reaction products susceptible to degradation by Uracil-DNA Glycosylase (UDG)—minimizes risk of carryover PCR-product contamination in subsequent reactions (UDG not included)
  • RNase inhibitor to protect RNA template from degradation
  • Additive that reduces primer-dimer formation, enhancing reaction specificity
  • Passive internal reference based on proprietary ROX™ dye for increased data precision

Minimal Nonspecific Amplification

Primer-dimer formation can be a significant issue for SYBR® Green-based qPCR. SYBR® Green Dye fluorescences when bound to any double-stranded DNA, including primer-dimers. If primer-dimers are present in your real-time PCR reaction, they can produce significant background fluorescence, confusing results. The Power SYBR® Green RNA-to-CT™ 1-Step Kit is designed to reduce primer-dimer formation and nonspecific amplification. Figure 1 shows the results when no-template control samples (NTC) are amplified using a set of 91 PCR primer pairs with the Power SYBR® Green RNA-to-CT™ 1-Step Kit, or with three other commercially available one-step qRT-PCR kits. The results show significant nonspecific signal in reactions that used other vendors’ kits whereas results from the reaction using the Power SYBR® Green RNA-to-CT™ 1-Step Kit shows low background signals.

 Figure 1: Dissociation curves of the NTC from a 91-Assay Panel using the Power SYBR® Green RNA-to-Ct™ 1-Step Kit, as compared to the One-Step RT-PCR kits from three commercially available vendors.

High Sensitivity and Confidence

The Power SYBR® Green RNA-to-CT™ 1-Step Kit formulation helps to enhance sensitivity while reducing nonspecific amplification signal.

The kit can detect GAPDH gene product in as little as 0.1 pg of total RNA. Coupled with its ability to minimize nonspecific amplification, the Power SYBR® Green RNA-to-CT™ 1-Step Kit gives you confidence in data quality.

 Figure 2: Power SYBR® Green RNA-to-CT™ 1-Step Kit delivers high sensitivity. GAPDH target amplified from 0.1 pg to 1 pg of FirstChoice® Human Total RNA Survey Panel in 19 replicate reactions using the Applied Biosystems 7500 Fast Real-Time PCR System.