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For decades, Invitrogen SuperScript Reverse Transcriptases (RT) have supported reliable cDNA synthesis across a wide range of RNA research applications. Successive generations of SuperScript RTs have advanced reverse transcription performance through improvements in thermostability, processivity, inhibitor tolerance, and reaction speed, helping researchers work with increasingly demanding RNA samples and workflows.
Invitrogen SuperScript V products build on the proven performance of SuperScript IV RT with next-generation reverse transcription performance and greater workflow flexibility. Available as a stand-alone reverse transcriptase and ready-to-use VILO Master Mix formats, the SuperScript V family combines robust cDNA synthesis with improved reagent stability, flexible formats, and room-temperature workflows.
Comparison of SuperScript Reverse Transcriptases
Table 1. Key features of four generations of SuperScript Reverse Transcriptases.
| SuperScript V RT | SuperScript IV RT | SuperScript III RT | SuperScript II RT | |
|---|---|---|---|---|
| Performance | ||||
| Sensitivity | 0.1 pg | 1–10 pg | 10 pg | 1 ng |
| Maximum cDNA length | Up to 20 kb | Up to 13 kb | Up to 12.3 kb | Up to 12.3 kb |
| Optimal RT temperature | 50–55°C | 50–55°C | 50°C | 42°C |
| Thermostability | Up to 65°C | Up to 65°C | Up to 55°C | Up to 50°C |
| Challenging RNA | High | High | Medium | Low |
| Inhibitor tolerance | High | High | Medium | Low |
| Ease of set up | ||||
| Reaction time | 10 min or less | 10 min | 30–50 min | 50 min |
| Reaction set up | Room temperature | Ice | Ice | Ice |
| Stability of assembled reactions | Up to 24 hours* | No | No | No |
| DTT required | No | Yes | Yes | Yes |
| Stability | ||||
| Shipping conditions | Ambient Dry / wet ice |
Dry ice | Dry ice | Wet ice |
| Store at room temperature | Yes* | No | No | No |
| Animal-origin free | Yes | No | Yes | Yes |
*Stand-alone enzymes can be stored up to 6 months. Master mixes can be stored up to 1 month.
Why choose SuperScript V RT?
- Choose SuperScript V RT for excellent performance across routine and challenging RNA samples, with easy reaction setup and exceptional stability.
- For SuperScript II or III users, SuperScript V products provide a path to improved performance and a more streamlined workflow.
- For SuperScript IV RT users, in addition to performance, SuperScript V products offer workflow and stability benefits.
Which format to choose from
 SuperScript V Reverse Transcriptase |
 SuperScript V VILO Master Mix |
 SuperScript V VILO Master Mix, Primer-Free |
|---|---|---|
Stand-alone RT for customized cDNA synthesis
|
Ready-to-use master mix with optimized oligo(dT) and random primers
|
Ready-to-use master mix without primers
|
|
||
Advantages of SuperScript V Reverse Transcriptase
Excellent performance
Reliable cDNA synthesis across a broad range of RNA inputs and sample types, including challenging RNA.
- Sensitivity
- Efficiency
- Robust
- Long cDNA
- Thermostability
Sensitivity—Broad RNA input range (Figure 1). Sensitive down to 0.1 pg of total RNA.
Figure 1. Ultra-broad dynamic range and sensitive detection in qPCR. Distinct, well separated amplification curves demonstrate consistent performance across multiple orders of magnitude.
Efficient—Lower Ct values in RT-qPCR compared to other RTs (Figures 2 & 3).
Figure 2. Earliest Ct values compared to 19 other commercial RT kits and master mixes. SuperScript V Reverse Transcriptase consistently achieves earlier Ct values (red line), reflecting higher cDNA yield and enhanced detection sensitivity. All products were evaluated using manufacturer-provided protocols.
Figure 3. Significant Ct delay observed with competing RT master mixes. Bars represent the Ct delay observed with competitor kits relative to SuperScript V VILO Master Mix, which showed earlier Ct values of up to 8 cycles for genes tested.
Robust—Strong performance with low-input, degraded, highly structured, and inhibitor-rich RNA (Figures 4 & 5).
Figure 4. Robust performance in the presence of common RT inhibitors. SuperScript V Reverse Transcriptase consistently delivers the earliest Ct values across all tested conditions, demonstrating strong resistance to inhibition.
Figure 5. Reliable performance with degraded RNA samples. Comparison of Ct values for SuperScript V VILO Master Mix and leading competitor RT kits using degraded REN RNA (RIN=3). The master mix consistently delivers the earliest Ct values, indicating efficient cDNA synthesis even from compromised RNA templates.
Long cDNA synthesis—Transcribes RNA templates up to 20 kb cDNA (Figure 6).
Figure 6. Long cDNA synthesis up to 20 kb. Agarose gel analysis of RT-PCR products generated from mouse heart RNA. SuperScript V products enable efficient synthesis and amplification of full-length cDNA approaching 20 kb.
Efficient transcription at elevated reaction temperatures up to 65°C.
Figure 7. SuperScript V RT synthesizes up to 9 kb cDNA.
Easy set up
Simplified cDNA synthesis with fast reactions, room temperature setup, and greater workflow flexibility including fewer steps and stability of assembled reactions.
- Fast
- Room temperature set up
- Flexible workflow timing
11-minute cDNA synthesis time. Incubation time can be tailored for transcript length. Short incubation time streamlines processing by eliminating primer annealing and reducing heat inactivation to 1 minute (Figures 8, 9 & 10).
Figure 8. Streamlined workflow reduces reverse transcription time and simplifies processing. 13-minute cDNA synthesis workflow with gDNA removal.
Figure 9. Rapid cDNA synthesis enables detection in as little as 30 seconds. Efficient cDNA synthesis was observed across all time points, with clear detection of a 0.5 kb fragment even after just 30 seconds of reverse transcription.
Figure 10. Reliable target detection with reduced reaction times. RT-qPCR analysis os FZD6 (low-abundance) targets using 10 ng total RNA demonstrates consistent cDNA synthesis across a broad temperature range (37–65°C) and reduced reverse transcription incubation times (1–10 minutes).
Reactions can be set up at room temperature, without ice buckets or cooling racks (Figure 11).
Figure 11. Reaction setup at room temperature vs. on a cooling rack (automation compatibility). SuperScript V VILO Master Mix reactions containing a TaqMan Gene Expression Assay targeting MYC were assembled either at room temperature or on a cooling rack and incubated for up to 45 minutes during reaction setup. Subsequent RT-qPCR analysis shows comparable Ct values between setup conditions.
Reactions assembled with SuperScript V VILO Master Mix remain stable for up to 24 hours at room temperature.
Figure 12. RT-PCR product yield remained consistent across a broad RNA input range when reactions were processed immediately or held at room temperature prior to reverse transcription.
Exceptional stability
Greater stability at ambient temperatures provides added flexibility for shipping and storage while reducing dependence on cold-chain conditions.
- Ambient shipping—Suitable for ambient shipping, simplifying logistics and reducing dependence on cold-chain transport (Figure 13 & 14).
- Room-temperature storage—SuperScript V Reverse Transcriptase can be stored at room temperature for up to 6 months. Master Mixes can be stored for up to 1 month.
Figure 13. SuperScript V VILO Master Mix maintains RT-qPCR performance after storing at room temperature for one month. Cₜ values remained consistent across high-, medium-, and low-abundance targets compared with frozen control.
Figure 14. SuperScript V enzyme activity remained stable after extended storage at room termperature.
SuperScript IV Reverse Transcriptases for specialty applications formats
- Standalone reverse transcriptase
- Ready-to-use master mix
- Primer free master mix
SuperScript V Reverse Transcriptases FAQs
Find tips, troubleshooting help, and resources for common questions about SuperScript V Reverse Transcriptases. Can’t find your question?
SuperScript V Reverse Transcriptase products support first-strand cDNA synthesis workflows, including RT-qPCR, RT-PCR, gene expression analysis, RNA sequencing library preparation, and analysis of low-input or challenging RNA samples.
Yes. SuperScript V Reverse Transcriptase products are designed to generate high-quality cDNA suitable for sensitive and reproducible qPCR analysis.
For maximal convenience, we recommend SuperScript V VILO Master mix format (Cat. No. 11800001) which provides reverse transcriptase, RNase inhibitor, buffer, MgCl₂, dNTPs, and an optimized blend of oligo (dT) and random primers in a single formulation, optimized for two-step RT-qPCR applications.
cDNA prepared using SuperScript V Reverse Transcriptase products is compatible with most commercially available qPCR master mixes. However, for optimal performance the following reagents are recommended:
- TaqMan system: TaqMan Fast Advanced qPCR Master Mix.
- SYBR Green system: PowerTrack SYBR Green qPCR Master Mix
SuperScript V Reverse Transcriptase products support reverse transcription in as little as 1–10 min depending on the workflow and target abundance. For most applications, we recommend a 10 min reverse transcription incubation step.
SuperScript V Reverse Transcriptase products include a room-temperature reaction control feature that helps suppress reverse transcription activity during reaction setup. This feature allows assembled reactions to remain stable at room temperature before incubation.
Yes. SuperScript V Reverse Transcriptase products are designed to support ambient shipping conditions, which may reduce reliance on dry ice or other cold-chain shipping methods.
Resources
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For Research Use Only. Not for use in diagnostic procedures.