Details
Masterful reverse transcriptase
- The Scriptase III is an enhanced variant of the Murine Leukemia Virus (MuLV) reverse transcriptase.
- Due to specific mutations in the RNase H domain of the reverse transcriptase, cDNA can be synthesized even from very small amounts of RNA.
- Scriptase III has additional mutations that make it even more robust and thermostable. It can therefore operate with maximum activity up to a temperature of 55 °C. At these elevated temperatures, highly structured RNAs can easily unfold, allowing them to serve as templates for reverse transcription. It is the superior choice for complex tasks, such as Real-Time PCR and NGS.
Improved engineered enzyme – Designed for extensively structured RNA
- FastGene® Scriptase III is an improved version of M-MLV Reverse Transcriptase, which exhibits high thermal stability (~55°C) and low RNase H activity.
- This enzyme is capable of synthesizing first strand cDNA at 42℃~55℃ with high specificity and yield. At such high temperatures, extensively structured RNAs can be readily unfolded to serve as templates for reverse transcription.
- SuperiorScript III can synthesise long cDNAs (up to ~12 kb).
Applications
- Quantification of gene Expression
- Real-Time PCR
- Next generation sequencing
- cDNA library construction
- Difficult templates with RNA secondary structures
- Very low RNA concentrations
Quality control
- Purity: >99% on SDS-PAGE
- Endonuclease- and exonuclease-free
- RNase-free
- Inhibitor-free
- Satisfactory cDNA product yield and length
Components
FastGene® Scriptase III comes with additional components that are needed for reverse transcription.
It consists of 5 vials including:
- FastGene® Scriptase III Restriction Enzyme
- 5X First-Strand Buffer (250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15mM MgCl2)
- 0.1 M DTT
- dNTP mix (10 mM each)
- Sterile water (RNase free)
This product requires the purchase of primers (oligo dT, random hexamer) separately.
The FastGene® Scriptase III is also available as cDNA Synthesis Kit (LS66) or cDNA 5x ReadyMix (LS67), see ordering information.
Boost your cDNA synthesis with thermostable precision
Scriptase III sets a new standard for reverse transcription with its enhanced thermostability, allowing efficient operation at temperatures up to ~55°C. This elevated temperature helps unwind RNA secondary structures, granting better accessibility and increasing the sensitivity of reverse transcription reactions. The result? Higher yields of full-length, high-quality cDNA.
Figure 1: Performance of the Scriptase III at different temperatures. The thermostability of the Scriptase III ensures a reliable operation up to ~55 °C.
Amplify productivity with superior cDNA yields
Boost your productivity with Scriptase III. Real-time PCR analysis reveals significantly lower Cq values, indicating its efficiency in generating abundant and high-quality cDNA even from minimal RNA input.
Figure 2: Productivity of Scriptase III. Scriptase III shows excellent productivity and cDNA yields for different RNA targets, compared to competitor enzyme.
Incredible sensitivity
Achieve precise cDNA synthesis even from minimal RNA samples with Scriptase III. This advanced reverse transcription enzyme ensures reliable detection down to 0.1 ng of total RNA, as demonstrated with HeLa RNA extracts. Real-time PCR results show consistent GAPDH amplification across RNA concentrations ranging from 100 ng to 0.1 ng, showing Scriptase III’s unparalleled sensitivity and efficiency. Perfect for low-input applications, Scriptase III delivers consistent, high-quality cDNA to support your most demanding research needs.
Figure 3: Sensitivity of Scriptase III. Scriptase III ensures reliable detection down to 0.1 ng of total RNA extracts.
High-speed cDNA synthesis
When time is critical, Scriptase III rises to the challenge. Its reaction time can be shortened to 30 minutes if needed, without sacrificing cDNA yield or performance. This flexibility makes it an excellent choice for high-quality results even under a tight schedule.
Figure 4: Reaction time or Scriptase III. Fast reaction times of Scriptase III allow a reduction of the incubation time down to 30 minutes.
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