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PRODUCT

High Sensitivity Reverse Transcriptase

RT optimized for low-copy and structured RNAs

High‑sensitivity reverse transcriptase designed for precise, low‑copy detection and structured RNAs. Excels in standard RT, RT‑qPCR, primer extension, and RACE with clean backgrounds and consistent yields.

  • Sensitive RT for low‑copy targets
  • Handles structured RNAs cleanly
  • Supports RT‑qPCR, RACE, primer extension
  • Consistent yields with clean backgrounds

Select a Size

High Sensitivity Reverse Transcriptase (400 U)

#EM16HSREVT-Sm
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Specifications Description Quality Resources

Product Specifications

Product Number EM16HSREVT
Intended Use This product is for research use only
Function Engineered RT optimized for specificity and sensitivity
For Use In RT-qPCR, low-copy RNA detection, RACE, primer extension
Concentration 4 U/µL
Source Recombinant E. coli
Shipping Conditions Cold Pack
Storage Temperature -20 C
Stability ≥12 months
Delivery Format Tubes
Description

High-Efficiency cDNA Synthesis

High Sensitivity Reverse Transcriptase is a proprietary enzyme designed for highly specific and sensitive reverse transcription, delivering exceptional performance across a wide range of RNA analysis applications. With its high affinity for RNA, it efficiently transcribes complex RNA secondary structures and targets in low copy numbers, resulting in high yields of cDNA.

This multifunctional enzyme combines RNA-dependent and ssDNA-dependent DNA polymerase activities with RNase H activity that is specific to RNA hybridized to cDNA. This specificity ensures no degradation of pure RNA templates, improving the performance of subsequent PCR steps. High Sensitivity Reverse Transcriptase is ideal for standard reverse transcription, RT-PCR, qRT-PCR, and advanced workflows like RACE and primer extension RNA analysis.

Key Features

  • High Sensitivity and Specificity: Guarantees accurate reverse transcription, even with low-copy targets.
  • Exceptional RNA Affinity: Transcribes complex RNA secondary structures with ease.
  • High cDNA Yields: Produces robust cDNA output, even from challenging templates.
  • RNase H Activity: Specific to RNA hybridized to cDNA, ensuring improved 1-step PCR performance.

Applications

  • Standard Reverse Transcription: Reliable synthesis of cDNA for downstream applications.
  • Synthesis of Double-Stranded cDNA: Ideal for cloning workflows.
  • RT-PCR and qRT-PCR: High sensitivity and specificity for RNA quantification.
  • Rapid Amplification of cDNA Ends (RACE): Perfect for studying RNA ends.
  • RNA Analysis by Primer Extension: Enables precise RNA structure and sequence analysis.

Contamination Prevention Tips

To ensure RNA integrity and prevent contamination during cDNA synthesis:

  • Use separate clean areas for sample preparation and reaction setup.
  • Use DEPC-treated tubes and pipette tips or certified nuclease-free labware.
  • Wear fresh gloves and handle RNA with care.
  • Assess RNA integrity using denaturing agarose gel electrophoresis before cDNA synthesis.
  • Use RNase-free water and reagents, and optionally add Ribonuclease inhibitor (20 units per 20 µl reaction).

Custom Options

We offer flexible customization to meet your specific needs:

  • Volume: Optimize packaging sizes to match your application scale.
  • Concentration: Customize concentrations for optimal performance.
  • Formulation: Available in liquid and lyophilized formats to suit your protocols.

Quality

One unit is the amount of enzyme activity that incorporates 1 nmol of dTTP into acid insoluble fraction in 10 minutes at 42°C when poly(A)+ RNA and oligo(dT)20 are used as template–primer.

Exonuclease assay
Linearized lambda/HindIII fragments are incubated with the Reverse Transcriptase in a 50 µl reaction mixture for 16 h at 37°C. No degradation of DNA was observed.

Endonuclease/Nick Activity
Supercoiled plasmid DNA is incubated with the enzyme in a 50 µl reaction mixture for 4 h at 37°C. No conversion of covalently closed circular DNA to nicked DNA detected.

Contamination with E. coli DNA
Absence of E. coli genomic DNA is confirmed by qPCR using a sample of the enzyme and specific primers targeting the E. coli 16S rRNA gene. No contamination detected.

RNase Assay
An RNA template is incubated with the enzyme in a 20 µl reaction mixture for 1 h at 42°C. No RNA degradation observed.

Functional Assay
cDNA synthesis with Oligo (dT) and/or Hexamer primers, followed by PCR.

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