Ultra-Clean UCF.ME™ Thermostable RNase H: Precise Cutting for More Efficient Experiments!

Ideal for rRNA removal and mRNA capping efficiency detection!

Thermostable RNase H, with its exceptional heat resistance, outperforms regular RNase H in both specificity and efficiency!  

Introduction to RNase H

E. coli RNase H (E. coli Ribonuclease H) is an enzyme that specifically degrades the RNA component of RNA-DNA hybrid strands. It plays a vital role in processes such as DNA replication, repair, and transcription by cleaving the RNA strand to maintain the integrity and stability of the DNA template. This makes it widely used in molecular biology experiments, including cDNA synthesis, rRNA removal, and RNA interference studies. However, E. coli RNase H has some limitations:

• It may cause non-specific cleavage of single-stranded RNA or DNA, reducing the accuracy of experimental results.
• Its poor thermal stability prevents it from maintaining activity at high temperatures, making it unsuitable for experiments like high-temperature reverse transcription or PCR that require elevated temperatures.

These shortcomings have driven researchers to develop thermostable RNase H to broaden its applications and enhance experimental efficiency.

Figure 1: RNase H Mechanism

Thermostable RNase H from Thermus thermophilus

Thermostable RNase H, derived from Thermus thermophilus, is a homolog of E. coli RNase H and shares similar ribonuclease functions. It precisely identifies and efficiently cleaves the phosphodiester bonds of the RNA strand in RNA:DNA hybrids while preserving the DNA strand’s integrity. In-depth structural analysis reveals that, although its overall stability distribution resembles that of E. coli RNase H, T. thermophilus RNase H exhibits significant improvements in both overall stability and local residue stability.

With an optimal activity temperature above 65°C, Thermostable RNase H delivers enhanced specificity and efficiency at higher reaction temperatures, minimizing non-specific cleavage. This property unlocks its vast potential in molecular biology experiments, including:

• rRNA removal
• mRNA capping rate detection
• Removal of mRNA poly(A) hybridized to poly(dT)
• Removal of mRNA during cDNA second-strand synthesis
• Improved amplification efficiency in high-temperature reverse transcription, isothermal amplification, and PCR experiments

Figure 2: Three-Dimensional Structure Comparison of Thermostable RNase H and E. coli RNase H ^[1]

 UCF.ME™Thermostable RNase H (Cat14545)

Thermostable RNase H is frequently used in pathogen detection experiments, such as rRNA removal in metagenomic sequencing (mNGS) and pathogen-targeted sequencing (tNGS). Residual host or background bacterial nucleic acids in the enzyme can significantly compromise detection accuracy. To address this, Yeasen Biotech introduces UCF.ME™ Thermostable RNase H (Cat14545), developed using its proprietary UCF.ME™ ultra-clean process technology. Produced at the UCF.ME™ ultra-clean molecular enzyme facility, this product undergoes rigorous quality control—from material selection and environmental management to process optimization and testing—ensuring minimal background bacterial gDNA and nuclease residues. This guarantees accurate, reliable, and reproducible experimental results.

Product Advantages:

• High activity and excellent batch-to-batch consistency
• Extremely low host gDNA residue: <0.02 Copies/U
• No exonuclease, endonuclease, or RNase residues
• Outstanding stability: No significant loss of enzyme activity after 32 days at 4°C, 16 days at 25°C, or 7 days at 37°C

Performance Showcase of UCF.ME™ Thermostable RNase H (Cat14545)

1. High Activity and Batch Consistency

Three batches of UCF.ME™ Thermostable RNase H were incubated with RNA:DNA substrates, and band changes were analyzed via agarose gel electrophoresis. Results demonstrate that just 0.05 U of this enzyme effectively cleaves the RNA in 20 pmol RNA:DNA substrates, with excellent batch-to-batch consistency, highlighting its stability and reliability.

Figure 3: Activity Detection Results of UCF.ME™ Thermostable RNase H  

Note: Reaction conditions: 50°C for 20 minutes; RNA:DNA substrate – 20 pmol

2. Low Host gDNA Residue: <0.02 Copies/U

Host (E. coli) gDNA residue testing across multiple batches shows that all three batches of UCF.ME™ Thermostable RNase H have residue levels well below 0.02 Copies/U, ensuring high purity and experimental reliability.

Figure 4: Host gDNA Residue Results of UCF.ME™ Thermostable RNase H

3. No Exonuclease, Endonuclease, or RNase Residues

25 U of UCF.ME ™ Thermostable RNase H was incubated with nucleic acid substrates, and band changes were assessed by agarose gel electrophoresis. No exonuclease, endonuclease, or RNase residues were detected in any of the three batches, providing strong assurance for result accuracy and reliability.

Figure 5: Detection Results of Exonuclease, Endonuclease, and RNase Residues in UCF.ME™ Thermostable RNase H

4. Excellent Stability

UCF.ME™ Thermostable RNase H was subjected to stability tests: 32 days at 4°C, 16 days at 25°C, and 7 days at 37°C. Enzyme activity measurements showed no significant decline, proving its exceptional stability across a wide temperature range and its suitability for long-term storage and diverse experimental conditions.

Figure 6: Accelerated Stability Results of UCF.ME™ Thermostable RNase H

 Recommended Related Products

Source	Product Name	Cat No.
T. thermophilus	UCF.ME™ Thermostable RNase H	14545ES
E. coli	RNase H	12906ES

 

 References

1. Hollien J, Marqusee S. Structural distribution of stability in a thermophilic enzyme [J]. Proceedings of the National Academy of Sciences, 1999, 96(24): 13674-13678.

2. Wolf EJ, Dai N, Chan SH. Selective Characterization of mRNA 5′End-Capping by DNA Probe-Directed Enrichment with Site-Specific Endoribonucleases [J]. [2025-03-03].

3. Gu H, Sun YH, Li XZ. Novel rRNA-depletion methods for total RNA sequencing and ribosome profiling developed for avian species [J]. Poultry Science, 2021, 100(7): 101321. DOI:10.1016/j.psj.2021.101321.