Thermo Scientific™ FastDigest™ BamHI

Thermo Scientific™ FastDigest BamHI is a high-efficiency restriction enzyme that recognizes and cleaves the palindromic DNA sequence 5'- G↓GATCC -3' / 3'- CCTAG↑G -5'. Designed for rapid DNA digestion in just 5–15 minutes, this enzyme is fully active in the universal FastDigest Buffer system, eliminating the need for buffer changes or additional DNA cleanup steps.

FastDigest BamHI provides 100% compatibility with single, double, and multi-enzyme digestions, ensuring high accuracy and specificity without star activity. It is widely used in molecular cloning, restriction mapping, genotyping, and other DNA modification techniques.

Key Features & Benefits

1. Ultra-Fast DNA Digestion in 5–15 Minutes
   • Reduces restriction digestion time without compromising specificity.
   • Enables efficient plasmid digestion for cloning and gene editing.
2. 100% Activity in Universal FastDigest Buffers
   • Eliminates the need for buffer changes and additional DNA cleanup.
   • Works seamlessly with FastDigest and FastDigest Green buffers.
3. No Star Activity for High-Fidelity Digestion
   • Ensures accurate cleavage with no off-target effects.
   • Maintains restriction site integrity in sensitive applications.
4. Direct Gel Loading with FastDigest Green Buffer
   • Allows immediate electrophoresis without additional preparation.
   • Includes tracking dyes and density reagents for streamlined workflow.
5. Compatible with DNA Modifying Enzymes
   • Supports direct addition of T4 DNA Ligase, Klenow Fragment, T4 DNA Polymerase, and Alkaline Phosphatase.
   • Ensures seamless transition from restriction digestion to DNA modification steps.

Product Specifications

Use Cases & Applications

1. Molecular Cloning & Recombinant DNA Technology

• Plasmid & Vector Digestion for Gene Cloning
  • Produces sticky ends for directional cloning.
  • Ensures efficient DNA fragment excision for subcloning applications.
• Double Digestion for Seamless DNA Assembly
  • Works well with other FastDigest enzymes, eliminating the need for multiple buffers.
  • Reduces reaction time while maintaining high restriction site specificity.

2. Genotyping & Genetic Variation Analysis

• Restriction Fragment Length Polymorphism (RFLP) Analysis
  • Differentiates genetic variants based on restriction enzyme digestion patterns.
  • Used for genotyping, marker-assisted selection, and forensic analysis.
• Single Nucleotide Polymorphism (SNP) Detection
  • Helps identify 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
  • Useful for epigenetic profiling and structural variation studies.
  • Helps differentiate methylation-sensitive DNA regions.

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 FastDigest BamHI 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 BamHI?

✅ 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 BamHI 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.