| Field | Specification |
|---|---|
| Product Type | |
| Shipping | |
| Source | Recombinant (E. coli) |
| Storage |
Bbs I is a Type IIS restriction endonuclease encoded by the BbsI gene of Brevibacillus laterosporus. FuniCut Series Endonucleases are fast restriction endonucleases that can accurately complete DNA cleavage within 5 to 15 minutes using genetic engineering recombination technology. They are suitable for rapid cleavage of plasmid DNA, PCR products, genomic DNA, etc. FuniCut rapid endonucleases share a common enzyme cleavage buffer, simplifying the enzyme reaction system. In addition, they have good enzyme activity redundancy and can easily handle the enzymatic cleavage of excess substrates or difficult templates.
Specifications
|
Cat.No. |
14227ES76/14227ES90/14227ES94/14227ES95 |
|
Size |
500 U/5000 U/20,000U/200,000U |
|
Restriction Enzyme Cut Site |
5'-GAAGAC(N)₂↓-3' |
|
Recommend reaction conditions |
1×FuniCut™ Buffer, Incubate at 37°C |
|
Enzyme activity |
20 U/μL |
|
Inactivation condition |
Incubate at 80℃ for 20 min |
|
Isoschizomer |
BstV2I, BpiI |
Glycerol Content |
Contains Glycerol |
Components
|
Components No. |
Name |
14227ES76 (500 U) |
14227ES94 (20KU) |
|
14227-A |
FuniCut™ Bbs I |
25 μL |
1 mL |
|
14227-B |
10×FuniCut™ Buffer |
1 mL |
3 x 1 mL |
|
14227-C |
10×FuniCut™ Color Buffer* |
1 mL |
- |
【Note】: *10×FuniCut Color Buffer contains red and yellow tracking dyes which allow the product to be used directly for gel electrophoresis. The red dye in FuniCut Color Buffer has a migration rate similar to a 2500 bp double-stranded DNA fragment in 1% agarose gel; the yellow dye has a migration rate similar to a 10 bp double-stranded DNA fragment in 1% agarose gel.
Storage
This product should be stored at -25~-15℃ for 2 years.
Notes
1. Star activity was not observed after 3 h incubation, but delayed enzyme digestion may result in star activity.
2. This product is for research use only.
3. Please operate with lab coats and disposable gloves，for your safety.
Instructions
1. DNA rapid enzyme digestion protocol
1) Prepare the system reaction solution using the following recommended sample addition sequence (operate on ice)
|
Components |
Plasmid DNA |
PCR products |
gDNA |
|
ddH2O |
15 μL |
16 μL |
30 μL |
|
10×FuniCut Buffer or 10×FuniCut Color Buffer |
2 μL |
3 μL* |
5 μL |
|
Substrate DNA |
2 μL (~1 μg) |
10 μL (~0.2 μg) |
10 μL (5 μg) |
|
FuniCut Bbs I |
1 μL |
1 μL |
5 μL |
|
Total |
20 μL |
30 μL |
50 μL |
【Note】：*This system refers to purified PCR products. Unpurified PCR products have a certain ionic strength and the amount of 10×FuniCut Buffer added can be reduced accordingly to 2 μL. If cloning and other experiments are performed in the next step, the PCR product must be purified before enzyme digestion.
2) Mix by gentle pipetting or swirling (do not vortex), then centrifuge briefly to collect the droplets.
3) Incubate at 37°C for 15 min (plasmid), 15-30 min (PCR product) or 30-60 min (genomic DNA).
4) Incubate at 80°C for 20 min to inactivate the enzyme and stop the reaction (optional).
5) If FuniCut Colour Buffer is used for the digestion reaction, the product can be loaded directly onto the electrophoresis.
2. Double or multiple enzyme digestion protocol
1) The dosage of each endonuclease is 1 µL and the reaction system should be expanded accordingly as needed.
2) The total volume of all endonucleases should not exceed 1/10 of the total reaction system.
3) If the optimal reaction temperatures of the selected endonucleases are different, use the enzyme with the lowest optimal temperature for enzyme digestion, then add the enzyme with the highest optimal temperature and incubate at a higher temperature.
3. Reaction system
|
Components |
Volume（20 μL） |
Volume（20 μL） |
Volume（50 μL）* |
|
DNA |
1 μg |
2 μg |
5 μg |
|
10×FuniCut Buffer or 10×FuniCut Color Buffer |
2 μL |
2 μL |
5 μL |
|
FuniCut Bbs I |
1 μL |
2 μL |
5 μL |
|
Total |
20 μL |
20 μL |
50 μL |
【Note】:*If the total reaction volume is greater than 20 μL, use a water, metal or sand bath and increase the incubation time.
4. Number of recognition sites in different DNA
|
λDNA |
ΦX174 |
pBR322 |
pUC57 |
pUC18/19 |
SV40 |
M13mp18/19 |
Adeno2 |
|
24 |
3 |
3 |
0 |
0 |
3 |
0 |
27 |
5. Effects of methylation modification
|
Dam |
Dcm |
CpG |
EcoKI |
EcoBI |
|
No influence |
No influence |
No influence |
No influence |
No influence |
6. Activity in different reaction buffers*
|
Reaction buffer |
FuniCut™ Buffer |
Thermo FastDigest Buffer |
NEB CutSmart™ Buffer |
Takara QuickCut™ Buffer |
|
Activity |
100% |
≤10% |
100% |
25% |
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 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 required to completely digest 1 µg of lambda DNA (or specified substrate DNA) in 60 min at the recommended incubation temperature in a total reaction volume of 50 µL.
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.
Restriction enzyme efficiency depends on buffer composition (NaCl/KCl concentration, pH, Mg²⁺), incubation temperature (typically 37°C), and template quality. Star activity—non-specific cleavage at relaxed recognition sequences—can occur under conditions of excess enzyme, extended incubation, high glycerol concentration, or non-optimal ionic strength. Use of manufacturer-supplied buffers and adherence to recommended enzyme:DNA ratios (typically 1–10 U per µg DNA) minimizes star activity.
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.
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