Thermo Scientific™ XbaI (10 U/μL)

Thermo Scientific™ XbaI is a high-quality restriction endonuclease that specifically recognizes and cleaves the palindromic DNA sequence 5'- T↓CTAGA -3' / 3'- AGATC↑T -5'. This enzyme is commonly used in molecular cloning, restriction mapping, genotyping, and genetic engineering applications.

XbaI performs efficient DNA cleavage at 37°C in Tango buffer, making it suitable for single and double digestion protocols, RFLP analysis, and Southern blotting. It is also available as a FastDigest enzyme for rapid DNA digestion.

Key Features & Benefits

1. High-Fidelity DNA Digestion
   • Recognizes T^CTAGA sites with consistent and efficient cleavage.
   • Suitable for vector cloning, site-directed mutagenesis, and DNA fragment isolation.
2. Optimized for Molecular Cloning & Genetic Engineering
   • Works best at 37°C in Tango buffer, ensuring efficient restriction digestion.
   • Included in Thermo Scientific Five Buffer System for universal enzyme compatibility.
3. Fast & Efficient DNA Cleavage
   • High activity (10 U/μL) for rapid DNA digestion.
   • Available as a FastDigest enzyme for streamlined workflows.
4. Dam Methylation-Sensitive for Selective Restriction Analysis
   • Sensitive to Dam methylation, meaning certain plasmid DNA preparations may be resistant to digestion.
   • Not sensitive to CpG or Dcm methylation, making it compatible with a broad range of genomic DNA sources.
5. Superior Quality & Purity
   • Free from nonspecific nuclease contamination.
   • Stringently tested to ensure reproducible and high-performance restriction digestion.

Product Specifications

Use Cases & Applications

1. Molecular Cloning & Recombinant DNA Technology

• Plasmid Digestion for Vector Cloning
  • Generates sticky ends for directional DNA ligation.
  • Enables precise DNA fragment insertion into expression vectors.
• Double Digestion for Efficient DNA Assembly
  • Works well with other Thermo Scientific restriction enzymes.
  • Compatible with FastDigest enzymes for rapid cloning workflows.

2. Genotyping & Genetic Variation Analysis

• Restriction Fragment Length Polymorphism (RFLP) Analysis
  • Used for identification of genetic markers and genotyping studies.
  • Differentiates DNA sequences based on XbaI restriction sites.
• Single Nucleotide Polymorphism (SNP) Detection
  • Identifies specific point mutations in genomic DNA.
  • Supports molecular diagnostic and genetic screening applications.

3. Southern Blotting & DNA Fragment Analysis

• Preparation of DNA Fragments for Probe Hybridization
  • Ensures specific DNA digestion for Southern blot applications.
  • Generates well-defined DNA fragments for genetic research.
• Chromosomal Analysis & Methylation Studies
  • Investigates epigenetic modifications using XbaI digestion patterns.
  • Helps differentiate methylation-sensitive DNA regions.

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

• Efficient Restriction Digestion for Library Preparation
  • Prepares DNA templates for sequencing applications.
  • Ensures uniform DNA fragment generation for high-quality sequencing data.
• Seamless Integration into Automated DNA Cloning Systems
  • Eliminates manual buffer changes using the Five Buffer System.
  • Reduces time and labor in restriction-based cloning protocols.

Recommended Protocol for XbaI 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 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 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 local biosafety regulations.
  • Use approved chemical waste disposal protocols for reaction waste.

Why Choose Thermo Scientific™ XbaI Restriction Enzyme?

✅ High specificity for T^CTAGA 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

✅ Dam methylation-sensitive enzyme for epigenetics and genotyping studies

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