DNA POLYMERASE I
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DNA Polymerase I
DNA Polymerase I is a mesophilic E. coli polymerase that catalyzes 5' → 3' template-directed DNA synthesis. DNA Polymerase I has 3' → 5' exonuclease (proofreading) activity and 5' → 3' exonuclease activity. It is capable of catalyzing de novo synthesis of synthetic homopolymers and provides a convenient method for the preparation of a variety of defined DNA substrates in the laboratory.
Key Features and Benefits
- Effective nick translation
- Reduce material costs with a drop-in solution that delivers equivalent performance at a differentiated price-point
- Custom formats, including high concentration, support lyophilization applications
- Highly stringent enzyme manufacturing ensures quality performance across lots
Applications
- Nick translation
- Second-strand synthesis
QC Specifications
| Description | Specification |
|---|---|
| Protein Purity | ≥ 99% |
| Endonuclease Contamination* | Not detectable |
| DNA contamination Assay ( E. coli , mammalian, library)* | < 10 copies |
| Phosphatase Contamination* | < 1% released |
| Unit Assay* | Pass |
*As assessed using 2 U of enzyme input per assay.
Properties
Unit definition: One unit is defined as the amount of DNA Polymerase I that will incorporate 300 nmol of dNTPs into a DNA template in 60 minutes at 37°C
Reaction conditions*:
- 1X DNAP I Reaction Buffer (not supplied with kit): 50 mM Tris-HCl, pH 7.9, 10 mM MgCl₂, 1 mM DTT
- Incubate at 37°C for ≥ 30 minutes
*Please reference our technical guide for instructions for use.
Storage buffer: 25 mM Tris-HCl, pH 7.4, 0.1 mM EDTA, 50% Glycerol, 1 mM DTT
Heat inactivation: 75°C for 20 min
Molecular weight: 103 kDa
5' – 3' Exonuclease activity: Yes
3' – 5' Exonuclease activity: Yes
Strand Displacement activity: No
Fidelity: High
Processivity: Moderate
BROCHURES
Precision Enzymes and Proteins Brochure
Custom Genomic Solutions Enzymes MDx DNA Polymerase I Equinox Library Amplification Equinox Uracil-Tolerant Library Amplification phi29 DNAP RNase Inhibitor RNase H StellarScript RT StellarScript HT RT StellarScript HT+ RT StellarTaq DNA Polymerase T4 DNA Ligase T4 DNAP T4 Polynucleotide Kinase T4 Gene 32 Protein Taq DNAPWatchmaker Genomics offers a portfolio of precision-engineered enzymes and proteins, designed to meet the demands of high-stringency applications. We provide tailored services such as custom fills, formats, packaging, and labeling to accommodate unique specifications.
PROTOCOLS
DNA Polymerase I Technical Guide
Enzymes DNA Polymerase IProtocol for Watchmaker Genomics' DNA Polymerase I Kit which enables high-fidelity DNA synthesis, second-strand synthesis, and nick translation. Ideal for defined DNA substrates; store at -20°C.
| CONFIGURATION | |
|---|---|
| Kit contents | DNA Polymerase I (10 U/µL) |
| Unit definition | One unit is defined as the amount of DNA Polymerase I that will incorporate 300 nmol of dNTPs into a DNA template in 60 minutes at 37°C |
| Storage buffer | 25 mM Tris-HCl, pH 7.4, 0.1 mM EDTA, 50% Glycerol, 1 mM DTT |
| QUALITY CONTROL | |
|---|---|
| Protein Purity | ≥ 99% |
| Endonuclease Contamination* | Not detectable |
| DNA contamination* (E. coli, mammalian, library) | <10 copies |
| Phosphatase Contamination* | <1% released |
| Unit Assay* | Pass |
*As assessed using 2 U of enzyme input per assay.
| SHIPPING AND HANDLING | |
|---|---|
| Shipping conditions | Ice packs |
| Storage | -20°C ± 5°C |
| Shelf life (off-the-shelf products) | ≥ 12 months |
| Shelf life (custom products) | ≥ 18 months |
What is DNA Polymerase I (DNAP I)?
DNA Polymerase I is a mesophilic E. coli polymerase that catalyzes 5'→3' template-directed DNA synthesis.
DNA polymerase I has 3'→5' exonuclease (proofreading) activity and 5'→3' exonuclease activity making it suitable for nick translation.
What is the unit definition of DNAP I?
One unit of DNA Polymerase I is defined as the amount of enzyme that will incorporate 300 nmol of dNTPs into a DNA template in 60 minutes at 37°C.
What are the relevant applications for DNAP I?
- Second-strand synthesis
- Nick translation
- Library preparation
What is the storage buffer for DNAP I?
Please see the Technical Guide for more information on DNA Polymerase I storage buffer.
Storage Buffer: 25 mM Tris-HCl, pH 7.4, 0.1 mM EDTA, 50% Glycerol, 1 mM DTT
What is the recommended reaction buffer for DNAP I?
DNA Polymerase I performs well in a range of buffers. The recommended 10X buffer composition is a standard reaction buffer. This buffer should be optimized for specific applications. The reaction buffer is not supplied with the enzyme. Please see the Technical Guide for more information on the recommended reaction buffer composition.
Recommended 10X DNA Polymerase I Reaction Buffer (not supplied with the kit): 500 mM Tris-HCl, pH 7.9 at 25°C, 100 mM MgCl₂, 10 mM DTT
Are reaction buffer and dNTPs supplied with the kit?
No, reaction buffer and dNTPs are not supplied with the enzyme.
What are the recommended reaction setup conditions?
Please reference our Technical Guide for instructions on recommended reaction setup conditions.
What is the optimal temperature for DNAP I?
The optimal temperature for DNA Polymerase I activity is 37°C.
How is DNAP I inactivated?
DNA Polymerase I is inactivated by heating at 75°C for 20 minutes, or through the addition of an equal volume of 0.25M EDTA pH 8.0.
Can DNAP I be used to blunt DNA?
Yes, DNA Polymerase I can be used to generate blunt ends. Its 3'→5' activity can remove 3′ overhangs and the polymerase activity will fill in 5' overhangs.
Can DNA Polymerase I be used to fill in/remove 3' overhangs and fill in/remove 5' overhangs?
Yes. DNA Polymerase I catalyzes DNA synthesis in the 5'→3' direction; its 3'→5' exonuclease activity removes 3' overhangs, its polymerase activity fills in 5' overhangs, and its 5'→3' exonuclease can remove 5' flaps or RNA primers—allowing it to both remove and fill overhangs in a single reaction to generate blunt ends.
What polymerase and exonuclease activity does DNA Polymerase I possess?
DNA Polymerase I catalyzes DNA synthesis in the 5'→3' direction, possesses both 3'→5' proofreading exonuclease activity and 5'→3' exonuclease activity (for nick translation and primer removal), and lacks strong strand‐displacement capability.
Does DNA Polymerase I exhibit strand displacement activity?
No, DNA Polymerase I lacks strand-displacement capability.
Can DNA Polymerase I amplify uracil-containing templates?
Yes, DNA Polymerase I can amplify uracil-containing templates.
Can DNA Polymerase I incorporate modified or alternative bases?
Yes, DNA Polymerase I can incorporate modified or alternative bases for DNA synthesis.
Where can I go for additional help or technical support?
If you need more detailed guidance or have questions not addressed here, please reach out to our support team at support@watchmakergenomics.com or submit a request via our Technical Support form. Please include details such as product name, lot number, version of protocol you’re using, and a brief description of your question or issue so we can assist you promptly.
Do you offer custom formats?
Yes! Watchmaker offers custom fill volumes, packaging, concentrations, and labeling – including private label – designed to meet your unique needs. We offer flexible terms to serve organizations of any size, and our processes enable rapid turnaround time on customization. Please contact sales@watchmakergenomics.com or submit a request via our Custom Solutions form to learn more about our capabilities.
Are you ISO 13485-certified?
Yes! Our Quality Management System has achieved ISO 13485:2016 Certification. Our certificate has been awarded for the design, development, manufacture, contract manufacture, and support of high performing reagents for genomics applications in medical research. Download the certificate here.
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| Description | 2.5 kU | |
|---|---|---|
| DNA Polymerase I (10 U/µL) | 7K0113-250UL | Request a quote |
Please contact sales@watchmakergenomics.com to inquire about custom kit configurations.
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