Bsu DNAP Highlights
Bsu DNA Polymerase, Large Fragment (Bsu) is a strand-displacing polymerase which retains the 5'→ 3' polymerase activity of the Bacillus subtilis DNA polymerase, but lacks the 5'→ 3' exonuclease domain. This large fragment lacks 3'→ 5' exonuclease activity. This polymerase — in combination with the T4 UvsX DNA Recombinase, T4 UvsY Protein, and T4 Gene 32 Protein — can be used to initiate polymerization via Recombinase Polymerase Amplification (RPA).
Bsu is available in glycerol-free and glycerol-containing formats.
- Individually QC’d for enzyme activity - ensuring highly reproducible performance
- Freeze-thaw stable formulation simplifies storage and handling
- Glycerol-free formulation available, suitable for lyophilization and microfluidic applications
Applications
- Recombinase Polymerase Amplification (RPA) for rapid, isothermal DNA amplification
- Isothermal amplification
- DNA strand displacement synthesis
- Second strand cDNA synthesis
- Random primer labelling
Properties
Unit definition: 1U of Bsu DNA Polymerase, Large Fragment is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 37°C.
Heat inactivation: 75°C for 20 min
Recommended reaction buffer (10x): 500 mM NaCl, 100 mM Tris-HCl, pH 7.9, 100 mM MgCl2, 10 mM DTT
5' – 3' Exonuclease activity: No
3' – 5' Exonuclease activity: No
Strand displacement activity: Yes
Key Performance Data
Consistent lot-to-lot activity
Enzyme and protein lot-to-lot variability has historically limited assay reliability – until now. In order to control concentration and activity of Bsu, an individual activity-based QC assay has been developed and implemented to ensure consistent enzyme production.
Figure 1. Consistent activity between lots ... MORE
Consistent lot-to-lot activity
Figure 1. Consistent activity between lots of Bsu DNA Polymerase, Large Fragment. Using in-house developed QC assays, enzyme activity levels were measured for multiple lots. Each lot of Bsu DNAP showed a high level of consistency, demonstrating robust production processes.
BROCHURES
Recombinase Polymerase Amplification Brochure
Custom Genomic Solutions Isothermal Amplification Pathogen Detection Enzymes MDx RNase H T4 Gene 32 Protein Bsu DNA Polymerase, Large Fragment Recombinase Polymerase Amplification (RPA) T4 UvsX DNA Recombinase T4 UvsY ProteinThis brochure presents the Watchmaker Recombinase Polymerase Amplification (RPA) enzymes including T4 UvsX DNA Recombinase, T4 UvsY Protein, T4 Gene 32 Protein, and Bsu DNA Polymerase, Large Fragment.
Custom Genomic Solutions Brochure
Custom Genomic Solutions Isothermal Amplification Pathogen Detection PCR/qPCR Rolling Circle Amplification RT-PCR/RT-qPCR Enzymes MDx 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 DNAP Bsu DNA Polymerase, Large Fragment Recombinase Polymerase Amplification (RPA) T4 UvsX DNA Recombinase T4 UvsY ProteinWhether you need small-scale modifications or a fully custom solution, Watchmaker’s flexible model ensures you get exactly what you need — with speed, simplicity, and scientific precision.
POSTERS
Optimization of Recombinase Polymerase Amplification (RPA) highlights the importance of enzyme consistency for reproducible results
Isothermal Amplification DNA Enzymes MDx T4 Gene 32 Protein Bsu DNA Polymerase, Large Fragment Recombinase Polymerase Amplification (RPA) T4 UvsX DNA Recombinase T4 UvsY ProteinThis poster presents data showing how Watchmaker individual enzyme QC for RPA enzymes leads to reliable, consistent amplification. Effects of damaged proteins in RPA reactions are measured, as well as the effect of multiple freeze-thaw cycles. As presented at ADLM 2025.
| CONFIGURATION | |
|---|---|
| Kit contents | Bsu DNA Polymerase, Large Fragment (50 U/μL) |
| Unit definition | 1U of Bsu DNA Polymerase, Large Fragment is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 37°C. |
| QUALITY CONTROL | |
|---|---|
| Purity (SDS-PAGE) | ≥ 95% |
| dsDNA exonuclease | < 1% released |
| ssDNA exonuclease | < 1% released |
| DNA contamination Assay (E. coli, mammalian, library)* | < 10 copies |
| Phosphatase Contamination Assay* | < 1% released |
| Endonuclease Contamination Assay* | Not detectable |
| Activity Assay | Passed |
* As assessed using 50 U or enzyme per assay.
| SHIPPING AND HANDLING | |
|---|---|
| Shipping | Glycerol-free: Dry Ice Glycerol-containing: Ice Packs |
| Storage | Glycerol-free: -80°C ± 10°C Glycerol-containing: -20°C ± 5°C |
| Shelf-life (off-the shelf products) | ≥ 12 months |
| Shelf-life (custom products) | ≥ 18 months |
What is Bsu DNA Polymerase, Large Fragment?
Bsu DNA Polymerase, Large Fragment is a strand-displacing polymerase which retains the 5'→ 3' polymerase activity of the Bacillus subtilis DNA polymerase, but lacks the 5'→ 3' exonuclease domain. This large fragment naturally lacks 3'→ 5' exonuclease activity.
Is Bsu DNA Polymerase, Large Fragment available in glycerol-free format?
Yes, Bsu DNA Polymerase, Large Fragment is available in both glycerol-containing and glycerol-free format, which is suitable for downstream lyophilization.
How is one unit of Bsu DNA Polymerase, Large Fragment defined?
One unit (1U) of Bsu DNA Polymerase, Large Fragment is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 37°C.
What are the relevant applications for Bsu DNA Polymerase, Large Fragment?
- Recombinase Polymerase Amplification (RPA)
- Isothermal amplification
- DNA strand displacement synthesis
- Second strand cDNA synthesis
- Random primer labelling
Can Bsu DNA Polymerase, Large Fragment be used to blunt DNA?
No, Bsu DNA Polymerase, Large Fragment lacks the 3'→ 5' exonuclease activity necessary to remove 3' additions and is therefore not suitable for generating blunt ends.
For blunting DNA, use the Watchmaker T4 DNA Polymerase.
What is the recommended reaction buffer for strand-displacement reactions when using Bsu DNA Polymerase, Large Fragment?
The reaction buffer is not supplied with the enzyme. Please contact Technical Support for more information on the recommended reaction buffer composition.
What components are required for Recombinase Polymerase Amplification (RPA)?
RPA requires a number of different components. Bsu DNA Polymerase, Large Fragment can be used for Recombinase Polymerase Amplification (RPA) of DNA when combined with glycerol-free Watchmaker Genomics reagents T4 UvsX DNA Recombinase (7K0124), T4 UvsY Protein (7K0126) and T4 Gene 32 Protein (7K0127). Contact Technical Support for other components required for RPA which should be sourced separately.>
How do I set up an initial RPA reaction with Bsu DNA Polymerase, Large Fragment?
For suggestions on how to set up an initial RPA reaction, please contact Technical Support.
How do I design primers for RPA?
The following characteristics are of critical importance when designing RPA primers:
- The optimal length for RPA primers is 30–35 nt. Primers less than 30 nt are not recommended.
- Primer sequences should have between 30–70% GC content and have no single or dinucleotide base repeats.
- Primer sequences should also be devoid of complementary sequences which promote secondary structure hairpin loops and prevent self-dimerization or primer-primer interactions.
- Amplicon sequences should have between 35–60% GC content, with an optimal length of 150–450 bp.
- If possible, the primer should end with a G or C on the 3' terminus.
What is the optimal reaction temperature for Bsu DNA Polymerase, Large Fragment?
The optimal reaction temperature for Bsu DNA Polymerase, Large Fragment is 37°C but can vary from 25°C to 50°C depending on the application.
How is Bsu DNA Polymerase, Large Fragment inactivated?
Bsu DNA Polymerase, Large Fragment is inactivated by Incubation at 75°C for 20 minutes.
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 customer fills, 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 right-sized processes enable rapid turnaround time on customization. Please contact sales@watchmakergenomics.com 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.
| Description | 1.25 kU | 25 kU | |
|---|---|---|---|
| Bsu DNA Polymerase, Large Fragment - Glycerol-free (50 U/μL) | 7K0128-25UL | 7K0128-500UL | Request a quote |
| Bsu DNA Polymerase, Large Fragment (50 U/μL) | 7K0129-25UL | 7K0129-500UL | Request a quote |
Please contact sales@watchmakergenomics.com to inquire about custom kit configurations.
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