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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
- 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.
- 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.
- Fast & Efficient DNA Restriction Digestion
- High activity (10 U/μL) for rapid digestion in standard protocols.
- Available as a FastDigest™ enzyme for faster reactions.
- Superior Quality & Purity
- Stringently tested for consistent performance in molecular cloning.
- Free from nonspecific nuclease contamination.
- 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
| Feature | Details |
|---|---|
| Product Name | Thermo Scientific™ XhoI Restriction Enzyme |
| Recognition Site | 5'- C↓TCGAG -3' / 3'- GAGC↑TC -5' |
| Catalog Numbers | ER0691 (2,000 units), ER0692 (5 × 2,000 units), ER0695 (5,000 units) |
| Enzyme Concentration | 10 U/μL |
| Optimal Temperature | 37°C |
| Compatible Buffer | 10X Buffer R |
| Methylation Sensitivity | Sensitive to CpG Methylation, Not Sensitive to Dam/Dcm Methylation |
| Heat Inactivation | Yes |
| Type IIS RE | No |
| Storage Conditions | -20°C |
| Product Type | Restriction Enzyme |
| Research Category | Traditional Cloning |
| Unit Size | Each |
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
- 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
- Incubation:
- Incubate at 37°C for 1 hour for complete digestion.
- Enzyme Inactivation:
- Heat inactivate at 65°C for 20 minutes, if necessary.
- 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.
Digestion of agarose-embedded DNA
User Guide: XhoI, 10 U/uL, 2000U
User Guide: XhoI, 10 U/uL, 5000U
User Guide: XhoI, 10 U/uL, 5x2000U
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
- 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.
- 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.
- Fast & Efficient DNA Restriction Digestion
- High activity (10 U/μL) for rapid digestion in standard protocols.
- Available as a FastDigest™ enzyme for faster reactions.
- Superior Quality & Purity
- Stringently tested for consistent performance in molecular cloning.
- Free from nonspecific nuclease contamination.
- 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
| Feature | Details |
|---|---|
| Product Name | Thermo Scientific™ XhoI Restriction Enzyme |
| Recognition Site | 5'- C↓TCGAG -3' / 3'- GAGC↑TC -5' |
| Catalog Numbers | ER0691 (2,000 units), ER0692 (5 × 2,000 units), ER0695 (5,000 units) |
| Enzyme Concentration | 10 U/μL |
| Optimal Temperature | 37°C |
| Compatible Buffer | 10X Buffer R |
| Methylation Sensitivity | Sensitive to CpG Methylation, Not Sensitive to Dam/Dcm Methylation |
| Heat Inactivation | Yes |
| Type IIS RE | No |
| Storage Conditions | -20°C |
| Product Type | Restriction Enzyme |
| Research Category | Traditional Cloning |
| Unit Size | Each |
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
- 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
- Incubation:
- Incubate at 37°C for 1 hour for complete digestion.
- Enzyme Inactivation:
- Heat inactivate at 65°C for 20 minutes, if necessary.
- 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.
Digestion of agarose-embedded DNA
User Guide: XhoI, 10 U/uL, 2000U
User Guide: XhoI, 10 U/uL, 5000U
User Guide: XhoI, 10 U/uL, 5x2000U
Specifications
| Pack Size | 2,000 UNITS, 5,000 units, 5 x 2,000 UNITS |