STELLARSCRIPT REVERSE TRANSCRIPTASE PORTFOLIO

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Precision Enzymes to fuel cutting-edge applications

The StellarScript® Reverse Transcriptase (RT) portfolio features three M-MLV variants with reduced RNase H activity and enhanced processivity, activity, thermostability, and inhibitor tolerance in comparison to wild-type enzyme. Kits contain all required reagents for cDNA synthesis except primers. The portfolio includes:

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Tailored reverse transcriptases to meet your application needs

  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.

Improve specificity with increased thermostability

The high degree of secondary structure in many RNA species, such as high-GC transcripts, can make full-length generation of cDNA difficult and decrease overall assay sensitivity. Increased reverse transcription temperatures relax secondary structure, but this approach requires a highly thermostable reverse transcriptase. The StellarScript portfolio contains variants with a wide range of optimal reverse transcription temperatures to increase cDNA product specificity.

Figure 1A. Overcome RNA template secondary structure with thermostable RTs. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT-primed first strand synthesis for 30 minutes using a 0.5 to 9 kb RNA ladder, as indicated. Reverse transcriptase thermostability increases from StellarScript to StellarScript HT and StellarScript HT+.
Figure 1B. Overcome RNA template secondary structure with thermostable RTs. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT-primed first strand synthesis at 42°C, 50°C, 60°C, or 65°C for 30 minutes using 10 ng total liver RNA as template. Resulting cDNA mass was assessed via qPCR. StellarScript and StellarScript HT activities drop at temperatures above 50°C, whereas StellarScript HT+ does not.
Activity at elevated temperatures
Figure 1A. Overcome RNA template secondary structure with thermostable RTs. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT-primed first strand synthesis for 30 minutes using a 0.5 to 9 kb RNA ladder, as indicated. Reverse transcriptase thermostability increases from StellarScript to StellarScript HT and StellarScript HT+.
Figure 1A. Overcome RNA template secondary structure with thermostable RTs. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT-primed first strand synthesis for 30 minutes using a 0.5 to 9 kb RNA ladder, as indicated. Reverse transcriptase thermostability increases from StellarScript to StellarScript HT and StellarScript HT+.
Figure 1B. Overcome RNA template secondary structure with thermostable RTs. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT-primed first strand synthesis at 42°C, 50°C, 60°C, or 65°C for 30 minutes using 10 ng total liver RNA as template. Resulting cDNA mass was assessed via qPCR. StellarScript and StellarScript HT activities drop at temperatures above 50°C, whereas StellarScript HT+ does not.
Figure 1B. Overcome RNA template secondary structure with thermostable RTs. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT-primed first strand synthesis at 42°C, 50°C, 60°C, or 65°C for 30 minutes using 10 ng total liver RNA as template. Resulting cDNA mass was assessed via qPCR. StellarScript and StellarScript HT activities drop at temperatures above 50°C, whereas StellarScript HT+ does not.
High-quality performance across a wide range of RNA inputs and qualities

The StellarScript portfolio delivers excellent performance across a wide range of RNA inputs and qualities, including highly damaged FFPE material and inputs as low as 10 pg. This provides broad application and sample type compatibility.

Figure 2A. Generate cDNA from low-input and degraded RNA samples. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT and randomly primed first strand synthesis at 42°C (StellarScript) or 50°C (HT and HT+) for 40 minutes using 100 ng to 0.01 ng total liver RNA as template. Resulting cDNA yields were assessed via qPCR. All enzymes show similar sensitivity when reverse transcription is employed in their optimal temperature range.
Figure 2B. Generate cDNA from low-input and degraded RNA samples. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT and randomly primed first strand synthesis at 50°C for 30 minutes using total RNA inputs from two independent FFPE blocks as indicated. Resulting cDNA yields were assessed via qPCR. All enzymes show similar sensitivity.
Reliable performance across sample inputs and qualities
Figure 2A. Generate cDNA from low-input and degraded RNA samples. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT and randomly primed first strand synthesis at 42°C (StellarScript) or 50°C (HT and HT+) for 40 minutes using 100 ng to 0.01 ng total liver RNA as template. Resulting cDNA yields were assessed via qPCR. All enzymes show similar sensitivity when reverse transcription is employed in their optimal temperature range.
Figure 2A. Generate cDNA from low-input and degraded RNA samples. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT and randomly primed first strand synthesis at 42°C (StellarScript) or 50°C (HT and HT+) for 40 minutes using 100 ng to 0.01 ng total liver RNA as template. Resulting cDNA yields were assessed via qPCR. All enzymes show similar sensitivity when reverse transcription is employed in their optimal temperature range.
Figure 2B. Generate cDNA from low-input and degraded RNA samples. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT and randomly primed first strand synthesis at 50°C for 30 minutes using total RNA inputs from two independent FFPE blocks as indicated. Resulting cDNA yields were assessed via qPCR. All enzymes show similar sensitivity.
Figure 2B. Generate cDNA from low-input and degraded RNA samples. StellarScript, StellarScript HT, and StellarScript HT+ were run in oligo-dT and randomly primed first strand synthesis at 50°C for 30 minutes using total RNA inputs from two independent FFPE blocks as indicated. Resulting cDNA yields were assessed via qPCR. All enzymes show similar sensitivity.
For assay developers – make realizing your next product easier
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Want more information? Visit our other StellarScript portfolio product pages:

Watchmaker StellarScript Reverse Transcriptase Banner

StellarScript

Robust cDNA synthesis from as little as 10 pg with template switching activity.

Watchmaker StellarScript HT Reverse Transcriptase Banner

StellarScript HT

Improved thermostability and inhibitor tolerance improves compatibility with challenging applications.

Watchmaker StellarScript HT+ Reverse Transcriptase Banner

Stellarscript HT+

Excellent thermostability and inhibitor tolerance, paired with template switching capability, delivers high-quality performance with a variety of samples, targets, and applications.

Coming soon: High-purity RNase Inhibitor

Protect what’s precious for your single cell, single nuclei, and RT-qPCR workflows.

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