Thermo Scientific™ FastDigest XbaI

Thermo Scientific™ FastDigest XbaI is a high-performance restriction enzyme that recognizes and cleaves the palindromic DNA sequence 5'- T↓CTAGA -3' / 3'- AGATC↑T -5'. This enzyme enables fast, high-fidelity digestion of DNA in just 5–15 minutes using the universal FastDigest Buffer system.

FastDigest XbaI is optimized for single, double, or multiple restriction digestion reactions and is fully compatible with downstream applications without requiring buffer changes or additional purification steps. The enzyme works efficiently at 37°C, with no star activity, ensuring reliable and reproducible restriction site cleavage for molecular cloning, genotyping, and genetic engineering.

Key Features & Benefits

1. Ultra-Fast DNA Digestion in 5–15 Minutes
   • Reduces restriction digestion time without compromising accuracy.
   • Ensures high efficiency in molecular cloning and genotyping workflows.
2. 100% Activity in Universal FastDigest Buffers
   • Eliminates the need for buffer changes or additional DNA cleanup steps.
   • Works seamlessly with FastDigest and FastDigest Green buffers.
3. No Star Activity for High-Fidelity Digestion
   • Maintains specificity with no unintended cleavage or over-digestion.
   • Produces consistent fragment sizes for downstream cloning applications.
4. Direct Gel Loading with FastDigest Green Buffer
   • Allows immediate electrophoresis analysis without additional preparation.
   • Includes tracking dyes and density reagents for simplified workflows.
5. Compatible with DNA Modifying Enzymes for Seamless Cloning
   • Supports direct addition of T4 DNA Ligase, Alkaline Phosphatase, Klenow Fragment, and T4 DNA Polymerase.
   • Enables efficient multi-step DNA modification workflows.

Product Specifications

Use Cases & Applications

1. Molecular Cloning & Recombinant DNA Technology

• Plasmid & Vector Digestion for Cloning
  • Produces sticky ends for precise vector-insert ligation.
  • Ensures accurate DNA fragment excision for subcloning experiments.
• Efficient 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 and marker-assisted selection in research studies.
• Single Nucleotide Polymorphism (SNP) Detection
  • Helps identify point mutations in genomic DNA samples.
  • Supports molecular diagnostics and genetic screening.

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
  • Detects methylation status in Dam-sensitive restriction sites.
  • Useful for epigenetic profiling and structural variation studies.

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 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 XbaI 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 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™ FastDigest XbaI?

✅ 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 XbaI 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.