Thermo Scientific™ EcoRI (10 U/μL)

Thermo Scientific™ EcoRI is a high-quality restriction enzyme that specifically recognizes and cleaves the palindromic DNA sequence 5'- G↓AATTC -3' / 3'- CTTAA↑G -5'. This enzyme is widely used in molecular cloning, restriction site mapping, genotyping, and Southern blot analysis.

EcoRI efficiently digests DNA at 37°C in its own optimized buffer, ensuring high specificity and consistent performance in restriction digestion experiments. It is also available in a FastDigest™ format for rapid DNA digestion.

Key Features & Benefits

1. High-Specificity Restriction Digestion
   • Recognizes and cleaves G^AATTC restriction sites with precision.
   • Produces sticky ends for efficient ligation and cloning applications.
2. Optimized for Molecular Cloning & Genetic Engineering
   • Works best at 37°C in its unique EcoRI buffer.
   • Part of Thermo Scientific’s Five Buffer System, enabling universal compatibility.
3. Efficient DNA Cleavage for Reliable Performance
   • High enzyme activity (10 U/μL) for rapid DNA digestion.
   • Also available as a FastDigest enzyme for accelerated workflows.
4. Compatible with Downstream Applications
   • Works seamlessly with DNA ligases, polymerases, and modifying enzymes.
   • Produces fragments ideal for recombinant DNA techniques.
5. Stable & High-Quality Manufacturing
   • Stringently tested to ensure high purity and consistency.
   • Free from nonspecific nuclease contamination.

Product Specifications

Use Cases & Applications

1. Molecular Cloning & Recombinant DNA Technology

• Plasmid & Vector Digestion for Gene Cloning
  • Produces sticky ends for directional ligation and cloning.
  • Ensures efficient DNA fragment excision for subcloning applications.
• Double Digestion for Seamless DNA Assembly
  • Works efficiently with other restriction enzymes for multi-fragment cloning.
  • Reduces reaction time and enhances accuracy in restriction-based cloning.

2. Genotyping & Genetic Variation Analysis

• Restriction Fragment Length Polymorphism (RFLP) Analysis
  • Identifies genetic variants based on restriction enzyme digestion patterns.
  • Used for genotyping, marker-assisted selection, and forensic analysis.
• Single Nucleotide Polymorphism (SNP) Detection
  • Helps detect point mutations and insertions/deletions in genomic DNA.
  • Supports molecular diagnostics and genetic screening applications.

3. Southern Blotting & DNA Fragment Analysis

• Preparation of DNA Fragments for Hybridization Assays
  • Ensures specific and reproducible restriction digestion for probe hybridization.
  • Used in gene mapping and epigenetic studies.
• Chromosomal Analysis & Methylation Studies
  • Suitable for methylation-dependent restriction analysis.
  • Helps in differentiating genetic variations and structural modifications.

4. High-Throughput Cloning & Next-Generation Sequencing (NGS)

• Efficient Restriction Digestion for Library Preparation
  • Generates consistent DNA fragments for sequencing library construction.
  • Ensures uniform fragment size for quality sequencing reads.
• Compatible with Automated Workflows & Liquid Handling Systems
  • Ideal for robotic sample processing in high-throughput genomic workflows.
  • Eliminates time-consuming buffer exchanges, streamlining DNA digestion workflows.

Recommended Protocol for EcoRI Digestion

Standard 50 µL Reaction Setup:

Incubation & Post-Digestion Processing

1. Digest at 37°C for 1 hour for complete restriction.
2. Heat inactivate at 65°C for 20 minutes, if necessary.
3. Analyze digestion products on a 1–2% agarose gel using ethidium bromide or SYBR Safe staining.

Storage & Handling Instructions

• Storage Conditions:
  • Store at -20°C for long-term enzyme stability.
  • Avoid freeze-thaw cycles to maintain enzyme activity.
• Handling Precautions:
  • Use sterile, nuclease-free reagents and pipette tips to prevent contamination.
  • Work in a DNA-free environment to ensure reproducibility.
• Disposal Guidelines:
  • Dispose of enzyme and buffers according to biosafety waste regulations.
  • Use approved chemical waste disposal protocols for reaction waste.

Why Choose Thermo Scientific™ EcoRI?

✅ High specificity for G^AATTC restriction sites with no star activity

✅ Optimized for molecular cloning, restriction mapping, and genetic engineering

✅ Compatible with the universal Tango buffer for double digestions

✅ Reliable and consistent performance in traditional and high-throughput workflows

✅ CpG methylation-sensitive enzyme for epigenetics and genotyping studies

Thermo Scientific™ EcoRI is a highly precise, fast-acting restriction enzyme, making it a trusted choice for molecular biology, cloning, and DNA analysis applications.