Thermo Scientific™ FastDigest™ EcoRI

Thermo Scientific™ FastDigest EcoRI is a high-fidelity restriction enzyme that recognizes and cleaves the palindromic DNA sequence 5'- G↓AATTC -3' / 3'- CTTAA↑G -5'. Designed for fast, high-precision DNA digestion within 5–15 minutes, this enzyme is 100% active in the universal FastDigest Buffer system, allowing for single, double, or multiple enzyme digestions without the need for buffer changes or DNA cleanup steps.

FastDigest EcoRI is fully compatible with downstream applications, including ligation, PCR, sequencing, and cloning, ensuring reliable and reproducible results in molecular cloning, restriction mapping, genotyping, and Southern blotting.

Key Features & Benefits

1. Ultra-Fast DNA Digestion (5–15 Minutes)
   • Enables rapid DNA cleavage with high specificity.
   • Saves valuable time in restriction digestion workflows.
2. 100% Activity in Universal FastDigest Buffers
   • Works seamlessly in FastDigest and FastDigest Green buffers.
   • Eliminates the need for buffer changes and extra purification steps.
3. No Star Activity for High-Fidelity Digestion
   • Ensures precise and consistent restriction site cleavage.
   • Prevents nonspecific DNA cleavage in molecular biology applications.
4. Direct Gel Loading with FastDigest Green Buffer
   • Allows immediate electrophoresis analysis without additional sample preparation.
   • Includes tracking dyes and density reagents for easy sample loading.
5. Compatible with DNA Modifying Enzymes for Seamless Cloning
   • Works alongside T4 DNA Ligase, Alkaline Phosphatase, Klenow Fragment, and T4 DNA Polymerase.
   • Ensures high-efficiency multi-step DNA modification workflows.

Product Specifications

Use Cases & Applications

1. Molecular Cloning & Recombinant DNA Technology

• Plasmid & Vector Digestion for Gene Cloning
  • Produces sticky ends for precise vector-insert ligation.
  • Ensures efficient DNA fragment excision for subcloning applications.
• Double Digestion for Seamless DNA Assembly
  • Compatible with other FastDigest enzymes, reducing reaction time.
  • Works efficiently in restriction-based cloning workflows.

2. Genotyping & Genetic Variation Analysis

• Restriction Fragment Length Polymorphism (RFLP) Analysis
  • Identifies genetic variations based on restriction site digestion.
  • Used in genotyping, genetic fingerprinting, and population studies.
• Single Nucleotide Polymorphism (SNP) Detection
  • Helps detect point mutations and small genetic variations.
  • Supports molecular diagnostics and genome-wide screening applications.

3. Southern Blotting & DNA Fragment Analysis

• Preparation of DNA Fragments for Hybridization Assays
  • Ensures accurate and reproducible restriction digestion for Southern blot analysis.
  • Useful in genomic mapping and methylation studies.
• Chromosomal Analysis & Methylation Studies
  • Detects methylation-sensitive restriction sites in genomic DNA.
  • Used for epigenetics and imprinting research.

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

• Efficient Restriction Digestion for DNA Library Preparation
  • Generates consistent DNA fragments for sequencing library construction.
  • Ensures uniform fragment size distribution for high-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 FastDigest EcoRI Digestion

Standard 50 µL Reaction Setup:

Incubation & Post-Digestion Processing

1. Digest at 37°C for 5–15 minutes for complete restriction.
2. Heat inactivate at 80°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™ FastDigest EcoRI?

✅ Ultra-fast digestion (5–15 minutes) for rapid cloning workflows

✅ 100% activity in universal FastDigest Buffers—no buffer changes needed

✅ No star activity—ensures high-fidelity restriction site cleavage

✅ Direct gel loading with FastDigest Green Buffer—eliminates extra steps

✅ Compatible with downstream applications, including PCR, ligation, and sequencing

Thermo Scientific™ FastDigest EcoRI is a high-efficiency restriction enzyme, enabling faster, more precise DNA digestion with reduced workflow complexity, making it ideal for cloning, sequencing, and molecular biology research.