Thermo Scientific™ RevertAid First Strand cDNA Synthesis Kit

The Thermo Scientific™ RevertAid First Strand cDNA Synthesis Kit provides a comprehensive system for efficient synthesis of first-strand cDNA from RNA templates. Designed for reverse transcription workflows, the kit includes all necessary components, including the RevertAid Reverse Transcriptase, RiboLock RNase Inhibitor, oligo(dT)18 primer, and random hexamer primers, enabling flexibility and optimal performance for a variety of RNA types.

Key Features:

1. Efficient Reverse Transcription:
   • Synthesizes full-length first-strand cDNA up to 13 kb, ideal for large RNA templates.
2. Broad Temperature Stability:
   • RevertAid Reverse Transcriptase is active at 42–50°C, ensuring optimal performance and minimizing RNA secondary structure issues.
3. RiboLock RNase Inhibitor:
   • Protects RNA from degradation, ensuring high-quality cDNA synthesis even at elevated temperatures.
4. Versatile Primer Options:
   • Includes oligo(dT)18 and random hexamer primers, enabling transcription of both poly(A)-tailed and non-poly(A)-tailed RNA.
5. Complete System:
   • All necessary reagents are included, eliminating the need for additional purchases.
6. Wide Application Range:
   • Suitable for RT-PCR, RT-qPCR, antisense RNA synthesis, and cDNA library construction.

Specifications:

Applications:

1. First-Strand cDNA Synthesis:
   • Ideal for RT-PCR and RT-qPCR workflows.
   • Enables analysis of gene expression, transcriptomics, and functional studies.
2. cDNA Library Construction:
   • Facilitates the creation of full-length cDNA libraries for downstream applications.
3. Antisense RNA Synthesis:
   • Supports the production of RNA probes for hybridization studies.
4. MicroRNA and Non-Poly(A) RNA Studies:
   • Random hexamer primers allow transcription of RNA species without poly(A) tails, such as microRNAs.

Protocol Overview:

1. Prepare Reaction Mix:
   • Combine template RNA, primers, and dNTP mix with the reaction buffer.
2. Incubate for Primer Annealing:
   • Heat the mixture to 65°C for 5 minutes, then chill on ice.
3. Add Reverse Transcriptase:
   • Add RevertAid Reverse Transcriptase and RiboLock RNase Inhibitor to the reaction.
4. Perform Reverse Transcription:
   • Incubate at 42°C for 60 minutes. For specific RNA templates, 50°C can be used.
5. Terminate Reaction:
   • Heat to 70°C for 5 minutes to terminate reverse transcription.
6. Use cDNA:
   • The synthesized cDNA is ready for downstream applications like PCR, qPCR, or library construction.

Kit Components:

• RevertAid Reverse Transcriptase
• RiboLock RNase Inhibitor
• 5X Reaction Buffer
• dNTP Mix
• Oligo(dT)18 Primer
• Random Hexamer Primer
• Control GAPDH RNA
• Forward and Reverse GAPDH Primers (10 µM)
• Nuclease-Free Water

Ordering Information:

Storage and Handling:

• Storage: Store at –20°C to maintain stability.
• Shipping: Shipped on dry ice to ensure product integrity.

Advantages:

1. High Yield and Efficiency:
   • Generates robust and full-length cDNA, even for large RNA templates.
2. Versatile Primer Compatibility:
   • Includes multiple primer options for various RNA templates and experiments.
3. Robust RNA Protection:
   • RiboLock RNase Inhibitor ensures RNA integrity during the process.
4. All-In-One Solution:
   • Contains all necessary components for reverse transcription.
5. Reliable and Reproducible:
   • Designed for high-quality results across a wide range of RNA sources.

Related Products:

• Thermo Scientific™ Maxima Reverse Transcriptase: For advanced reverse transcription workflows.
• Thermo Scientific™ Nuclease-Free Water: Ideal for RNA preparation and reaction setup.
• Thermo Scientific™ SYBR Green PCR Master Mix: For RT-qPCR applications.

Regulatory Notes:

• For Research Use Only: Not intended for diagnostic or therapeutic purposes.

The Thermo Scientific™ RevertAid First Strand cDNA Synthesis Kit is a versatile, reliable, and complete solution for efficient reverse transcription workflows. Its high performance and broad applicability make it an essential tool for RNA analysis, gene expression studies, and molecular biology research.