STELLARSCRIPT HT REVERSE TRANSCRIPTASE KITS

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StellarScript HT Highlights

  • Increased thermostability enhances cDNA generation from RNA with a high degree of secondary structure
  • Resistance to common inhibitors provides compatibility with a variety of sample sources and applications
  • Improved activity enables reverse transcription from input amounts as low as 10 pg
  • Kits contain all required reagents for cDNA synthesis, except primers, for simple experimental setup
  • Custom formats available, including those to support lyophilization applications
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Applications

  • First strand cDNA synthesis
  • Reverse transcription quantitative PCR
    (RT-qPCR)
  • Reverse transcription PCR (RT-PCR)
  • Primer extension
  • RNA sequencing (RNA-seq)
  • 3’ Rapid amplification of cDNA ends (RACE)
  • Applications leveraging SuperScript III for reverse transcription

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High-purity RNase Inhibitor

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Key Performance Data

Overcome secondary structure with a thermostable RT

It can be difficult to generate full-length cDNA from RNAs with a high degree of secondary structure, such GC-rich and viral RNAs. Reverse transcribing at elevated temperatures can relax this secondary structure, as well as reduce nonspecific primer hybridization.

StellarScript® HT generates higher cDNA yields and performs equivalently with respect to processivity relative to SuperScript III at elevated temperatures.

For maximum thermostability, see StellarScript HT+.

Figure 1A. Improved performance at elevated temperatures. Watchmaker’s StellarScript HT and ThermoFisher Scientific’s SuperScript III were run in oligo-dT-primed first strand synthesis at 42°C, 50°C, 60°C or 65°C for 25 minutes using 10 ng total liver RNA as template. Resulting cDNA mass was assessed via qPCR. StellarScript HT produced higher yields (indicated by lower Cq values) at any evaluated temperature than SuperScript III.
Figure 1B. Improved performance at elevated temperatures. Watchmaker’s StellarScript HT and ThermoFisher Scientific’s SuperScript III were run in oligo-dT-primed first strand synthesis at 42°C, 50°C or 55°C for 30 minutes using a 0.5 to 9 kb RNA ladder as template. Both enzymes have robust processivity across the temperatures assessed.
Activity at elevated temperatures
Figure 1A. Improved performance at elevated temperatures. Watchmaker’s StellarScript HT and ThermoFisher Scientific’s SuperScript III were run in oligo-dT-primed first strand synthesis at 42°C, 50°C, 60°C or 65°C for 25 minutes using 10 ng total liver RNA as template. Resulting cDNA mass was assessed via qPCR. StellarScript HT produced higher yields (indicated by lower Cq values) at any evaluated temperature than SuperScript III.
Figure 1A. Improved performance at elevated temperatures. Watchmaker’s StellarScript HT and ThermoFisher Scientific’s SuperScript III were run in oligo-dT-primed first strand synthesis at 42°C, 50°C, 60°C or 65°C for 25 minutes using 10 ng total liver RNA as template. Resulting cDNA mass was assessed via qPCR. StellarScript HT produced higher yields (indicated by lower Cq values) at any evaluated temperature than SuperScript III.
Figure 1B. Improved performance at elevated temperatures. Watchmaker’s StellarScript HT and ThermoFisher Scientific’s SuperScript III were run in oligo-dT-primed first strand synthesis at 42°C, 50°C or 55°C for 30 minutes using a 0.5 to 9 kb RNA ladder as template. Both enzymes have robust processivity across the temperatures assessed.
Figure 1B. Improved performance at elevated temperatures. Watchmaker’s StellarScript HT and ThermoFisher Scientific’s SuperScript III were run in oligo-dT-primed first strand synthesis at 42°C, 50°C or 55°C for 30 minutes using a 0.5 to 9 kb RNA ladder as template. Both enzymes have robust processivity across the temperatures assessed.
Increased resistance to common inhibitors

Inhibitors reduce cDNA synthesis efficiency and can adversely impact assay sensitivity. Improved inhibitor tolerance safeguards performance and provides compatibility across a wide range of sample types and sources.

StellarScript HT delivers superior inhibitor tolerance in comparison to StellarScript. For maximum inhibitor tolerance, see StellarScript HT+.

Figure 2. Reliable performance in the presence of common inhibitors. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT-primed first strand synthesis for 30 minutes using 10 ng total liver RNA as template in varying concentrations of common inhibitors. Resulting cDNA yields were assessed via qPCR. StellarScript HT provides enhanced inhibitor tolerance in comparison to StellarScript.
Increased resistance to common inhibitors
Figure 2. Reliable performance in the presence of common inhibitors. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT-primed first strand synthesis for 30 minutes using 10 ng total liver RNA as template in varying concentrations of common inhibitors. Resulting cDNA yields were assessed via qPCR. StellarScript HT provides enhanced inhibitor tolerance in comparison to StellarScript.
Figure 2. Reliable performance in the presence of common inhibitors. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT-primed first strand synthesis for 30 minutes using 10 ng total liver RNA as template in varying concentrations of common inhibitors. Resulting cDNA yields were assessed via qPCR. StellarScript HT provides enhanced inhibitor tolerance in comparison to StellarScript.
Generate cDNA from vanishingly low RNA inputs

StellarScript HT robustly synthesizes cDNA from a broad range of RNA inputs, including as little as 10 pg. This enhances compatibility with applications where RNA template is limiting.

Figure 3. StellarScript HT provides equivalent performance to SuperScript III. Watchmaker’s StellarScript HT and ThermoFisher Scientific’s SuperScript III were run in oligo-dT and randomly primed first strand synthesis at 50°C for 40 minutes using 1000 ng to 0.01 ng total liver RNA as template. Resulting cDNA yields were assessed via qPCR. StellarScript HT showed overall equivalency to SuperScript III.
Robust performance across inputs
Figure 3. StellarScript HT provides equivalent performance to SuperScript III. Watchmaker’s StellarScript HT and ThermoFisher Scientific’s SuperScript III were run in oligo-dT and randomly primed first strand synthesis at 50°C for 40 minutes using 1000 ng to 0.01 ng total liver RNA as template. Resulting cDNA yields were assessed via qPCR. StellarScript HT showed overall equivalency to SuperScript III.
Figure 3. StellarScript HT provides equivalent performance to SuperScript III. Watchmaker’s StellarScript HT and ThermoFisher Scientific’s SuperScript III were run in oligo-dT and randomly primed first strand synthesis at 50°C for 40 minutes using 1000 ng to 0.01 ng total liver RNA as template. Resulting cDNA yields were assessed via qPCR. StellarScript HT showed overall equivalency to SuperScript III.
For assay developers – make realizing your next product easier
  • Navigate the complexities of productization with an experienced team who’s as committed to your success as you are
  • All aspects of our customization process are designed to serve you with speed, agility, and above all else, a commitment to quality
  • Tailored fill volumes, labeling (including white and private label), and packaging designed to your specifications
  • All products are manufactured within an ISO 13485:2016-certified QMS (download our certificate)

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Property Application
Optimal
Temp		 Thermal
Stability		 Inhibitor
Tolerance		 Template
Switching		 RT-qPCR/
qRT-PCR		 RT-PCR RNA-seq scRNA-seq 5′ RACE 3′ RACE
StellarScript HT+ 42 – 65˚C +++ +++ + +++ +++ + + + +
StellarScript HT 42 – 50˚C ++ ++ – ++ ++ + – – +
StellarScript 42˚C + + + + + + + + +

Table 1. Overview of StellarScript portfolio enzyme properties and relevant applications. “+” signs indicate strength of a property or fit of an application for a specific enzyme.

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