Thermo Scientific™ XhoI (10 U/μL)

Thermo Scientific™ XhoI is a high-quality restriction endonuclease that recognizes and cleaves the palindromic DNA sequence 5'- C↓TCGAG -3' / 3'- GAGC↑T C -5'. This enzyme is commonly used in molecular cloning, restriction site mapping, genotyping, and Southern blotting applications.

XhoI performs efficient and precise DNA cleavage at 37°C in R buffer, making it ideal for high-fidelity restriction digestion, single and double digestion, and restriction fragment length polymorphism (RFLP) analysis. It is part of the Thermo Scientific Five Buffer System, ensuring 100% activity under optimized reaction conditions.

Key Features & Benefits

1. High-Quality Restriction Enzyme for Molecular Biology
   • Recognizes C^TCGAG sites with consistent and efficient cleavage.
   • Suitable for plasmid digestion, gene cloning, and DNA fragment excision.
2. Optimized for Reliable DNA Digestion
   • Works best at 37°C in R buffer, ensuring efficient DNA cleavage.
   • Compatible with the universal Tango buffer for double digestion experiments.
3. Fast & Efficient DNA Restriction Digestion
   • High activity (10 U/μL) for rapid digestion in standard protocols.
   • Available as a FastDigest™ enzyme for faster reactions.
4. Superior Quality & Purity
   • Stringently tested for consistent performance in molecular cloning.
   • Free from nonspecific nuclease contamination.
5. Methylation Sensitivity
   • Sensitive to CpG methylation, allowing for differential digestion of methylated and unmethylated DNA.
   • Not sensitive to Dam or Dcm methylation, ensuring compatibility with a wide range of plasmids.

Product Specifications

Use Cases & Applications

1. Molecular Cloning & Genetic Engineering

• Precise DNA Fragmentation for Cloning Experiments
  • Facilitates vector and insert preparation for recombinant DNA construction.
  • Used for sticky-end ligation in gene cloning workflows.
• Restriction Site Mapping & DNA Fragment Analysis
  • Identifies restriction sites in plasmid and genomic DNA.
  • Supports structural and sequence analysis of genetic elements.

2. Genotyping & SNP Analysis

• Genotyping & Mutation Detection
  • Enables single nucleotide polymorphism (SNP) and genetic variant analysis.
  • Supports differential digestion based on CpG methylation sensitivity.
• RFLP (Restriction Fragment Length Polymorphism) Analysis
  • Used in population genetics and genetic fingerprinting studies.
  • Helps detect mutations and genetic linkage markers.

3. Southern Blotting & Hybridization Studies

• DNA Fragmentation for Southern Blot Probing
  • Generates specific restriction fragments for hybridization assays.
  • Used in genomic DNA analysis and gene expression studies.
• Chromosomal & Epigenetic Analysis
  • Detects methylation status and structural variations in genomic DNA.
  • Useful in studies of epigenetic modifications and imprinting.

4. Double Digestion & Multi-Enzyme Reactions

• Efficient Double Digestion for Cloning & DNA Analysis
  • Works well with other restriction enzymes in the universal Tango buffer.
  • Reduces digestion time and improves reaction efficiency.
• Compatible with FastDigest™ Enzymes for Speed Optimization
  • Enables rapid DNA digestion for high-throughput molecular biology workflows.

Recommended Protocol for XhoI Digestion

1. Reaction Setup (Standard 50 µL Reaction Volume):
   • Plasmid/Genomic DNA: 0.5–1 µg
   • XhoI Enzyme (10 U/μL): 1 μL
   • 10X Buffer R: 5 μL
   • Nuclease-Free Water: Adjust to 50 μL
2. Incubation:
   • Incubate at 37°C for 1 hour for complete digestion.
3. Enzyme Inactivation:
   • Heat inactivate at 65°C for 20 minutes, if necessary.
4. Analysis of Digestion Products:
   • Perform agarose gel electrophoresis (1-2%) and visualize under UV light.

Storage & Handling Instructions

• Storage Conditions:
  • Store at -20°C to maintain enzyme stability.
  • Avoid freeze-thaw cycles to prevent enzyme degradation.
• Handling Precautions:
  • Use sterile, nuclease-free reagents to prevent contamination.
  • Always work in a clean, DNA-free environment.
• Disposal Guidelines:
  • Dispose of used enzyme and buffers according to biosafety waste regulations.

Why Choose Thermo Scientific™ XhoI Restriction Enzyme?

✅ High-fidelity restriction enzyme for precise DNA cleavage

✅ Compatible with Thermo Scientific’s Five Buffer System & Tango Buffer

✅ Validated for molecular cloning, RFLP, and Southern blot applications

✅ CpG methylation-sensitive enzyme for epigenetic and genotyping studies

✅ Optimized for single and double digestions in research and biotechnology

Thermo Scientific™ XhoI is a trusted, high-quality restriction enzyme, ensuring reliable performance in genetic engineering, molecular biology, and epigenetic studies.