STELLARSCRIPT HT+ REVERSE TRANSCRIPTASE KITS

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StellarScript HT+ Highlights

  • Excellent thermostability improves reverse transcription through RNA secondary structure
  • Robust inhibitor tolerance safeguards performance with a broad range of sample sources
  • Terminal transferase (TdT) activity delivers compatibility with template switching applications
  • 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
    (RT-qPCR)
  • Reverse transcription PCR (RT-PCR)
  • Primer extension
  • RNA sequencing (RNA-seq)
  • Single cell RNA sequencing (scRNA-seq)
  • 3’ and 5’ Rapid amplification of cDNA ends (RACE)
  • Several applications leveraging SuperScript IV and Maxima H Minus for reverse transcription

Key Performance Data

Improve performance with challenging RNA sequences

Increased thermostability enables reverse transcription at elevated temperatures to overcome RNA template secondary structure and generate cDNAs from difficult targets, such as viral RNA. It further improves specificity by minimizing nonspecific primer binding.

StellarScript HT+ is the most thermostable enzyme in the portfolio, generating higher cDNA yields and better maintaining its folded protein structure at elevated temperatures relative to SuperScript™ IV and Maxima H Minus.

Figure 1A. Excellent performance at elevated temperatures. Watchmaker’s StellarScript HT+ and ThermoFisher Scientific’s SuperScript IV and Maxima H Minus were run in oligo-dT-primed first strand synthesis for 25 minutes using 10 ng total liver RNA, as indicated. Resulting cDNA mass was assessed via qPCR. StellarScript HT+ produced higher yields (indicated by lower Cq values) at any evaluated temperature than SuperScript IV and Maxima H Minus, indicating improved efficiency at elevated temperatures.
Figure 1B. Excellent performance at elevated temperatures. Enzymes were further assessed via static light scattering while increasing the temperature to determine their respective aggregation temperatures, the point at which protein unfolding begins. The enhanced thermostability of StellarScript HT+ is further demonstrated with its increased aggregation temperature.
Figure 1C. Excellent performance at elevated temperatures. Watchmaker’s StellarScript HT+ and ThermoFisher Scientific’s SuperScript IV and Maxima H Minus were run in oligo-dT-primed first strand synthesis at 50°C, 55°C, 60°C or 65°C for 30 minutes using a 0.5 to 9 kb RNA ladder as template. All three enzymes have robust processivity up to 60°C.
Relax RNA secondary structure with increased reaction temperatures
Figure 1A. Excellent performance at elevated temperatures. Watchmaker’s StellarScript HT+ and ThermoFisher Scientific’s SuperScript IV and Maxima H Minus were run in oligo-dT-primed first strand synthesis for 25 minutes using 10 ng total liver RNA, as indicated. Resulting cDNA mass was assessed via qPCR. StellarScript HT+ produced higher yields (indicated by lower Cq values) at any evaluated temperature than SuperScript IV and Maxima H Minus, indicating improved efficiency at elevated temperatures.
Figure 1A. Excellent performance at elevated temperatures. Watchmaker’s StellarScript HT+ and ThermoFisher Scientific’s SuperScript IV and Maxima H Minus were run in oligo-dT-primed first strand synthesis for 25 minutes using 10 ng total liver RNA, as indicated. Resulting cDNA mass was assessed via qPCR. StellarScript HT+ produced higher yields (indicated by lower Cq values) at any evaluated temperature than SuperScript IV and Maxima H Minus, indicating improved efficiency at elevated temperatures.
Figure 1B. Excellent performance at elevated temperatures. Enzymes were further assessed via static light scattering while increasing the temperature to determine their respective aggregation temperatures, the point at which protein unfolding begins. The enhanced thermostability of StellarScript HT+ is further demonstrated with its increased aggregation temperature.
Figure 1B. Excellent performance at elevated temperatures. Enzymes were further assessed via static light scattering while increasing the temperature to determine their respective aggregation temperatures, the point at which protein unfolding begins. The enhanced thermostability of StellarScript HT+ is further demonstrated with its increased aggregation temperature.
Figure 1C. Excellent performance at elevated temperatures. Watchmaker’s StellarScript HT+ and ThermoFisher Scientific’s SuperScript IV and Maxima H Minus were run in oligo-dT-primed first strand synthesis at 50°C, 55°C, 60°C or 65°C for 30 minutes using a 0.5 to 9 kb RNA ladder as template. All three enzymes have robust processivity up to 60°C.
Figure 1C. Excellent performance at elevated temperatures. Watchmaker’s StellarScript HT+ and ThermoFisher Scientific’s SuperScript IV and Maxima H Minus were run in oligo-dT-primed first strand synthesis at 50°C, 55°C, 60°C or 65°C for 30 minutes using a 0.5 to 9 kb RNA ladder as template. All three enzymes have robust processivity up to 60°C.
Safeguard performance with robust inhibitor tolerance

Inhibitors can dampen reverse transcription efficiency and impact sensitivity in applications such as RT-qPCR. Improved robustness in the presence of common inhibitors safeguards performance and ensures compatibility across a wide range of sample types and sources.

StellarScript HT+ delivers the highest level of inhibitor tolerance across the StellarScript portfolio.

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+ outperforms both StellarScript and StellarScript HT.
Safeguard performance with robust inhibitor tolerance
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+ outperforms both StellarScript and 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+ outperforms both StellarScript and StellarScript HT.
Efficiently synthesize cDNA over a wide RNA input range

StellarScript HT+ delivers sensitive performance across a wide range of input amounts, including RNA inputs as low as 10 pg. This increases compatibility with template-limiting applications and can improve the achievable limit of detection. 

Figure 3. Generate cDNA from vanishingly low RNA inputs. Watchmaker’s StellarScript HT+ and ThermoFisher Scientific’s SuperScript IV and Maxima H Minus 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 IV and Maxima H Minus.
Efficiently synthesize cDNA over a wide RNA input range
Figure 3. Generate cDNA from vanishingly low RNA inputs. Watchmaker’s StellarScript HT+ and ThermoFisher Scientific’s SuperScript IV and Maxima H Minus 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 IV and Maxima H Minus.
Figure 3. Generate cDNA from vanishingly low RNA inputs. Watchmaker’s StellarScript HT+ and ThermoFisher Scientific’s SuperScript IV and Maxima H Minus 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 IV and Maxima H Minus.

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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 42˚C + + + + + + + + +
StellarScript HT 42 – 50˚C ++ ++ – ++ ++ + – – +
StellarScript HT+ 42 – 65˚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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