Thermo Scientific™ DNA Polymerase I (10 U/μL)

Thermo Scientific™ DNA Polymerase I is a highly versatile, template-dependent DNA polymerase that catalyzes 5'→3' synthesis of DNA while simultaneously exhibiting 3'→5' exonuclease (proofreading) and 5'→3' exonuclease activities. Additionally, it possesses ribonuclease H activity, making it an essential enzyme for a wide range of molecular biology applications.

Key Features:

1. Multifunctional Enzyme:
   • Performs 5'→3' DNA synthesis, 3'→5' proofreading, and 5'→3' exonuclease activities.
   • Includes ribonuclease H activity, enabling RNA-DNA hybrid processing.
2. Incorporates Modified Nucleotides:
   • Supports incorporation of biotin, digoxigenin, aminoallyl, and fluorescently-labeled nucleotides for advanced labeling applications.
3. Compatibility with Multiple Buffers:
   • Functions effectively in restriction enzyme, PCR, and RT buffers, offering flexibility in experimental workflows.
4. High Purity:
   • Manufactured under strict quality controls to ensure enzyme activity and reproducibility.

Applications:

1. DNA Labeling by Nick Translation:
   • Used alongside DNase I for labeling DNA with modified nucleotides for hybridization and imaging studies.
2. Second-Strand cDNA Synthesis:
   • Essential for complementary DNA (cDNA) synthesis in conjunction with RNase H, particularly in reverse transcription workflows.
3. Molecular Biology Research:
   • Ideal for DNA repair, recombination studies, and advanced cloning techniques.
4. RNA-DNA Hybrid Processing:
   • Utilized in workflows requiring ribonuclease H activity for RNA degradation in RNA-DNA hybrids.

Specifications:

Protocol Overview:

1. Prepare Reaction Mix:
   • Combine DNA Polymerase I, DNA template, primer, and dNTPs in the recommended buffer.
2. Set Incubation Parameters:
   • Incubate at optimal temperature (typically 37°C) for the required time, depending on the application.
3. Terminate Reaction:
   • Stop the reaction using appropriate buffers or by heating.
4. Downstream Applications:
   • Use the synthesized or labeled DNA for hybridization, amplification, or analysis.

Ordering Information:

Storage and Handling:

• Storage Conditions: Store at -20°C to maintain enzyme activity.
• Shelf Life: Refer to product label for expiration details.

Advantages:

1. Versatile Functionality:
   • Supports a wide range of applications, including DNA synthesis, labeling, and cDNA generation.
2. High Efficiency:
   • Reliable incorporation of modified nucleotides enhances workflow flexibility.
3. Robust Compatibility:
   • Performs effectively in various reaction buffers, reducing the need for buffer optimization.
4. Validated for Research:
   • Manufactured under stringent quality controls for reproducibility and consistency.

Related Products:

• Thermo Scientific™ dNTP Mix: For efficient DNA synthesis.
• Thermo Scientific™ RNase H: For second-strand cDNA synthesis workflows.
• Thermo Scientific™ Nick Translation Buffer: Optimized for DNA labeling applications.

Regulatory Notes:

• For Research Use Only: Not for use in diagnostic or therapeutic applications.

The Thermo Scientific™ DNA Polymerase I (10 U/μL) provides a high-quality solution for efficient DNA synthesis, labeling, and molecular biology applications. Its multifunctionality and flexibility make it a trusted tool for researchers worldwide.