Description
Hifair™ Super Reverse Transcriptase is a next-generation reverse transcriptase, which was engineered from M-MLV. This enzyme lacks RNase H activity and exhibits significantly improved thermal stability, tolerating reaction temperatures of up to 70°C. It is highly suitable for efficient reverse transcription of RNA templates with complex secondary structures.
Applications
This product is suitable for RT-PCR, RT-qPCR, RT-LAMP, RNA-seq, and cDNA synthesis experiments. The reverse transcription reaction temperature can reach up to 70°C.
Unit Definition
One unit (U) of enzyme activity is defined as the amount of enzyme required to incorporate 1 nmol of dTTP into acid-insoluble material within 10 minutes at 37°C, using Poly(A).Oligo(dT) as the template-primer.
Component
|
Component |
Name |
14612ES92 |
14612ES93 |
|
14612 |
Hifair™ Super Reverse Transcriptase (200 U/μL) |
50 μL |
250 μL |
Storage Conditions
Store at -25˚C to -15˚C for up to 18 months.
Usage Instructions
First-Strand cDNA Synthesis Procedure
1. RNA Denaturation (Optional Step)
1. RNA denaturation helps to open secondary structures, significantly improving first-strand cDNA yield.
|
Component |
Volume |
|
RNase-free ddH2O |
to 13 μL |
|
Oligo(dT)18 (50 μM) or Random Primers (50 μM) or Gene-Specific Primers (2 μM) |
1 μL |
|
Template RNA |
Total RNA: 10 pg - 5 μg or mRNA: 10 pg - 500 ng |
· Heat at 65°C for 5 minutes, then quickly chill on ice for 2 minutes.
· Briefly centrifuge to collect the reaction liquid and add the reverse transcription reaction mix as shown below.
2. Reverse Transcription Reaction Setup (20 μL System)
|
Component |
Volume |
|
Reaction mixture from the previous step |
13 μL |
|
5× Reaction Buffer |
4 μL |
|
dNTP Mix (10 mM) |
1 μL |
|
Hifair® Super Reverse Transcriptase (200 U/μL) |
1 μL |
|
RNase Inhibitor (40 U/μL) |
1 μL |
3. Reverse Transcription Program Settings
|
Temperature |
Time |
|
25°C* |
5 min |
|
50-70°C |
15-30 min |
|
85°C** |
5 min |
*When using Random Primers, incubate at 25°C for 5 minutes. If using Oligo(dT)18 or Gene-Specific Primers, this step can be omitted.
85°C heating for 5 minutes inactivates the reverse transcriptase and can be adjusted as needed.
The reverse transcription product can be used immediately for subsequent PCR or qPCR reactions, stored short-term at -20°C, or for long-term storage at -80°C after aliquoting to avoid freeze-thaw cycles.
Precautions
1. Maintain a clean laboratory environment; wear clean gloves and masks. Ensure all experimental consumables are RNase-free to prevent contamination.
2. Perform all operations on ice to prevent RNA degradation.
3. Use high-quality RNA samples to ensure efficient reverse transcription.
4. This product is for research use only.
5. For your safety and health, wear a lab coat and disposable gloves while handling the product.
Documents:
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FAQ
The product is for research purposes only and is not intended for therapeutic or diagnostic use in humans or animals. Products and content are protected by patents, trademarks, and copyrights owned by Yeasen Biotechnology. Trademark symbols indicate the country of origin, not necessarily registration in all regions.
Certain applications may require additional third-party intellectual property rights.
Yeasen is dedicated to ethical science, believing our research should address critical questions while ensuring safety and ethical standards.