| Field | Specification |
|---|---|
| Mfr No | |
| Product Type | |
| Shipping | |
| Source | Recombinant (E. coli) |
| Storage |
Nb.BbvCI is a nicking endonuclease that cleaves only one strand of a dsDNA substrate. It introduces a nick on dsDNA substrates without cleaving the double helix, commonly used in isothermal amplification techniques such as SDA and RCA. The nick generated by Nb.BbvCI creates a DNA gap, initiating the strand-displacement activity of DNA polymerase, and enabling repeated cycles of nicking, displacement, and extension, thereby achieving exponential amplification of nucleic acids.
Specifications
|
Enzyme |
Nb.BbvCI |
|
Cat.NO. |
14232ES75 / 14232ES92 |
|
Size |
300 U / 10000 U |
|
Recognition Site |
5'-CCTCA GC-3' 3'-GGAGT↑CG-5' |
|
Enzyme Activity |
10 U/μL |
|
Recommended Reaction Conditions |
1× Nb.BbvCI Buffer; incubate at 37°C. |
|
Inactivation Conditions |
Incubate at 80°C for 20 minutes. |
Glycerol Content |
Contains Glycerol |
Components
|
Components No. |
Name |
14232ES75 |
14232ES92 |
|
14232-A |
Nb.BbvCI(10 U/μL) |
30 μL |
1 mL |
|
14232-B |
10× FuniCut™ Buffer |
1 mL |
6 mL |
|
14232-C |
10× FuniCut™ Color Buffer |
1 mL |
6 mL |
Shipping and Storage
This product should be stored at -25℃~-15℃ for up to 2 years.
Unit Definition
One unit (U) is defined as the amount of enzyme required to completely convert 1 µg of supercoiled p615 DNA into the open circular form in 50 µl of reaction mixture within 1 hour at 37°C.
Notes
1. Do not exceed 10% of the total reaction volume when adding enzyme to avoid star activity caused by excessive glycerol.
2. Additives in restriction enzyme storage buffers (e.g., salts, EDTA, or ethanol) may inhibit the reaction. The smaller the reaction volume, the stronger the inhibitory effect, especially when contaminants (e.g., glycerol, salts) from the enzyme storage buffer mix with the substrate solution.
3. For your safety and health, wear a lab coat and disposable gloves.
4. This product is intended for research use only.
Instructions
1. DNA Digestion Protocol
1.1 Prepare the reaction mixture on ice using the following recommended order:
|
Reagent |
Volume |
|
ddH₂O |
to 50 µl |
|
10× CutOneTM Buffer or 10× CutOneTM Color Buffer |
5 µl |
|
Substrate DNAa |
1 µg |
|
Nb.BbvCI (10 U/µl) |
1 µl |
|
Total |
50 µl |
[Note]:a. Phenol, chloroform, ethanol, EDTA, detergents, or high salt concentrations in the DNA substrate may inhibit Nb.BbvCI activity.
1.2 Mix gently by pipetting or tapping the tube (do not vortex), then briefly centrifuge to collect droplets on the tube wall.
1.3 Incubate at 37°C for 15 min to 3 h.
1.4 Stop the reaction by incubating at 80°C for 20 min to inactivate the enzyme, or alternatively, terminate the reaction by column purification or phenol/chloroform extraction.
2.Number of Nb.BbvCI Recognition Sites in Different DNA Substrates
|
DNA Substrate |
λDNA |
ΦX174 |
pBR322 |
pUC57 |
pUC18/19 |
SV40 |
M13mp18/19 |
Adeno2 |
|
Number of Sites |
7 |
3 |
0 |
0 |
0 |
0 |
2 |
9 |
3. Effect of Methylation Modifications
|
Methylation Type |
Dam |
Dcm |
EcoKI |
EcoBI |
|
Effect on Nb.BbvCI Activity |
No effect |
No effect |
No effect |
Cleavage blocked |
Store this enzyme at -20°C and avoid repeated freeze-thaw cycles to preserve catalytic activity. The product is shipped Dry Ice and remains stable for up to one year from the date of manufacture when stored under recommended conditions. Aliquoting the stock solution into single-use volumes is recommended for enzymes used infrequently to minimize thermal cycling of the bulk stock.
This enzyme is validated for use in Cloning & Assembly, NGS Library Preparation applications. It is suitable for use in both standard laboratory research settings and, where applicable, optimized reaction workflows requiring high specificity, sensitivity, or throughput.
One unit (U) is defined as the amount of enzyme that generates a nick in 1 µg of specified double-stranded DNA in 60 min at the recommended temperature, producing greater than 90% nicked circular product.
This enzyme is produced as Recombinant (E. coli) and supplied as a Research Use Only (RUO) reagent. Each lot is subjected to activity assay, purity assessment by SDS-PAGE, and functional validation prior to release. A Certificate of Analysis (CoA) and Safety Data Sheet (SDS) are available on request.
This enzyme is produced recombinantly in E. coli under stringent quality controls and validated on multiple substrate types. Key performance parameters include specific activity, substrate specificity, cofactor requirements, optimal pH/temperature range, and compatibility with downstream assays. Refer to the product datasheet for validated lot-specific activity data and recommended reaction conditions.
Yeasen Biotechnology supports custom enzyme solutions across multiple service lines — from GMP-grade bulk supply to directed enzyme engineering. Contact BioHippo to discuss requirements and initiate a project inquiry.
▶ GMP-Grade & Bulk Supply
Select Yeasen enzymes are available in GMP grade, manufactured in an ISO 13485-certified UCF.ME™ ultra-clean molecular enzyme facility with FDA Drug Master File (DMF) support.
- GMP-grade release testing and CoA documentation
- ISO 13485-certified production facility
- Scalable from milligram to multi-gram quantities
- Consistent lot-to-lot activity specifications
▶ Glycerol-Free & Custom Formulation
Glycerol-free enzyme formats are available for applications requiring lyophilization compatibility, liquid handling automation, or direct IVD master mix integration.
- Glycerol-free liquid format (standard and custom buffers)
- Lyophilization-ready enzyme preparation
- Custom reaction buffer optimization for specific assay conditions
- Compatible with freeze-drying workflows for point-of-care formats
▶ Molecular IVD RDC Service
Yeasen's Research and Development Contracting (RDC) team delivers end-to-end solutions for molecular diagnostic product development, covering enzyme selection through clinical validation support.
- Enzyme selection and performance matching
- Primer/probe design and reaction buffer optimization
- Sensitivity, specificity, and precision validation studies
- Stability studies and SNP evaluation
- Instrument platform compatibility assessment
▶ ZymeEditor™ Enzyme Engineering
Yeasen's proprietary ZymeEditor™ directed evolution and rational design platform enables the development of custom enzyme variants with tailored performance characteristics not available in off-the-shelf products.
- Directed evolution for enhanced thermostability, processivity, or fidelity
- Rational design for altered substrate specificity or cofactor requirements
- Library screening from Yeasen's proprietary enzyme variant collection
- Scale-up to commercial quantities upon candidate confirmation
ⓘ Customization services are fulfilled by Yeasen Biotechnology. Lead times and minimum order quantities vary by service type. Contact BioHippo for project scoping and pricing.
