Thermo Scientific™ FastDigest MspI

The Thermo Scientific™ FastDigest MspI is an advanced restriction enzyme designed for efficient and rapid DNA digestion. This enzyme belongs to the FastDigest™ line and is optimized to provide complete digestion in just 5–15 minutes at 37°C. Its universal buffer system and high compatibility with downstream applications make it a valuable tool in molecular biology workflows.

Recognition Site and Cleavage Pattern

• Recognition Sequence:
  • 5' C↓CGG 3'
  • 3' GGC↑C 5'
• Cleavage Details:
  • Cuts the DNA to produce 5' protruding ends, which are suitable for applications like molecular cloning.

Buffer Compatibility and Reaction Conditions

1. Universal FastDigest Buffer System:
   • All FastDigest enzymes work in the same universal buffer system, eliminating the need for buffer changes.
   • Enables seamless integration of additional enzymes for downstream processing, such as:
     • Klenow Fragment
     • T4 DNA Ligase
     • Alkaline Phosphatase
     • T4 DNA Polymerase
   • Supports direct addition of enzymes for post-restriction modifications.
2. Reaction Buffers:
   • 10X FastDigest Buffer (colorless): Ideal for applications requiring fluorescence analysis or UV concentration measurements.
   • 10X FastDigest Green Buffer: Contains tracking dyes and a density reagent for direct gel loading without additional preparation.
3. No Star Activity:
   • Ensures specific cleavage at the recognition site without unwanted cuts, even under prolonged incubation or high enzyme concentrations.
4. Heat Inactivation:
   • Not required, saving time and reducing workflow steps.
5. Reaction Conditions:
   • Optimal temperature: 37°C.
   • Complete digestion within 5–15 minutes.

Applications

The FastDigest MspI enzyme is suitable for a variety of molecular biology applications, including:

• Molecular Cloning: Precise and efficient DNA manipulation for creating recombinant constructs.
• Restriction Site Mapping: Identifying restriction sites in DNA for genetic studies.
• Genotyping: Determining genetic variations using restriction patterns.
• Southern Blot Analysis: Detecting specific DNA sequences.
• Restriction Fragment Length Polymorphism (RFLP): Studying genetic diversity or mutations.
• Single Nucleotide Polymorphism (SNP) Analysis: Detecting SNPs for genotyping or functional studies.

Key Features

1. Speed and Efficiency:
   • Fast digestion, eliminating the need for long incubation times.
2. Versatile Buffer System:
   • Allows for single, double, or multiple DNA digestions in one reaction without buffer changes.
3. Convenience:
   • The Green Buffer simplifies workflows with direct gel-loading capabilities.
4. Comprehensive Performance:
   • 100% enzyme activity across all FastDigest buffers, ensuring consistent results.
   • Compatibility with downstream modifying enzymes for DNA processing.
5. Extensive Product Range:
   • Part of a collection of 176 FastDigest enzymes, ensuring availability for diverse research needs.
6. Robust Design:
   • No star activity even under suboptimal conditions.
   • Works with both colorless and Green buffers for specific use cases.

Specifications

• Methylation Sensitivity:
  • Not sensitive to Dam, Dcm, or CpG methylation, ensuring consistent cleavage across a wide range of DNA samples.
• Reaction Temperature:
  • Optimal at 37°C, standard for most restriction enzymes.
• Enzyme Type:
  • Produces 5' protruding ends for flexible downstream applications.
• Quantity:
  • 400 μL enzyme, sufficient for 400 reactions.
• Buffers:
  • Includes 10X FastDigest Buffer (colorless) and 10X FastDigest Green Buffer for application-specific needs.
• Storage Conditions:
  • Store at –20°C to maintain enzyme stability and activity.

Product Package

• FastDigest MspI: 400 μL enzyme.
• 10X FastDigest Buffer: 1.0 mL.
• 10X FastDigest Green Buffer: 1.0 mL.

Notes and Considerations

• Green Buffer vs. Colorless Buffer:
  • The Green Buffer is optimized for direct gel loading, making it ideal for electrophoresis workflows.
  • Use the colorless buffer for applications involving fluorescence or UV analysis.
• Downstream Enzyme Compatibility:
  • DNA modifying enzymes retain full activity in the FastDigest Buffer, allowing seamless transitions between restriction and modification steps.
• For Research Use Only:
  • The product is not intended for diagnostic procedures.

This enzyme is an advanced, fast, and reliable tool for restriction digestion and cloning applications, offering high flexibility and efficiency in molecular biology workflows.