Thermo Scientific™ Eco31I (BsaI) (10 U/μL)

Thermo Scientific™ Eco31I (BsaI) is a Type IIS restriction enzyme that recognizes the asymmetric sequence 5'- GGTCTC(N1/5)↓ -3' / 3'- CCAGAG(N5)↑ -5' and cleaves outside its recognition site, generating 4-base overhangs. This property makes it highly valuable for Golden Gate Assembly, scarless cloning, and synthetic biology applications.

Eco31I (BsaI) efficiently digests DNA at 37°C in Buffer G and is optimized for single, double, and multiplex restriction digestion reactions. It is fully compatible with Thermo Scientific’s Five Buffer System, ensuring high specificity, no star activity, and seamless integration into molecular cloning workflows.

Key Features & Benefits

1. Type IIS Restriction Enzyme for Precision Cloning
   • Cleaves outside its recognition site, producing customizable 4-base overhangs.
   • Ideal for scarless cloning and Golden Gate Assembly.
2. Optimized for Fast & Efficient Digestion
   • Completes digestion in 5–15 minutes at 37°C.
   • Works best in Buffer G and is compatible with universal Tango buffer for double digestions.
3. No Star Activity for High-Specificity Digestion
   • Ensures precise and reproducible restriction site cleavage.
   • Maintains DNA integrity for downstream applications.
4. Direct Gel Loading with FastDigest Green Buffer
   • Enables immediate analysis without additional sample preparation.
   • Includes tracking dyes and density reagents for simplified workflow.
5. Compatible with Downstream DNA Modification Enzymes
   • Works seamlessly with T4 DNA Ligase, Alkaline Phosphatase, Klenow Fragment, and T4 DNA Polymerase.
   • Ensures smooth transition from digestion to DNA ligation and cloning.

Product Specifications

Use Cases & Applications

1. Molecular Cloning & Recombinant DNA Technology

• Golden Gate Assembly for Seamless DNA Construction
  • Produces customizable overhangs for seamless multi-fragment assembly.
  • Used in synthetic biology and modular DNA assembly.
• Restriction Site-Free (Scarless) Cloning
  • Ensures scarless DNA fragment assembly without additional bases.
  • Supports ligation-independent cloning methods.

2. Genotyping & Genetic Variation Analysis

• Restriction Fragment Length Polymorphism (RFLP) Analysis
  • Identifies genetic variations based on restriction site digestion.
  • Used in genotyping, forensic analysis, and marker-assisted selection.
• Single Nucleotide Polymorphism (SNP) Detection
  • Helps detect 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 Southern blotting.
  • Useful for gene mapping and epigenetic studies.
• Chromosomal Analysis & Methylation Studies
  • Detects methylation-sensitive restriction sites in genomic DNA.
  • Used for epigenetics and imprinting research.

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 high-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 Eco31I (BsaI) 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™ Eco31I (BsaI)?

✅ Type IIS restriction enzyme with flexible overhangs for modular DNA assembly

✅ Optimized for Golden Gate Assembly & synthetic biology applications

✅ Compatible with universal FastDigest Buffers for rapid digestion

✅ Efficient 5–15 minute digestion for high-throughput workflows

✅ Precise and reproducible cleavage for molecular cloning and gene editing

Thermo Scientific™ Eco31I (BsaI) is a high-precision, fast-acting restriction enzyme, making it a trusted choice for molecular biology, synthetic biology, and DNA engineering applications.