Introduction
"Gentaur Molecular" operates five major technical platforms concurrently, supporting a cutting-edge special enzyme research and development as well as production system. We offer:
- A complete range of mRNA raw materials and various mRNA stock solutions, continually optimizing and enhancing our products, with key quality indicators of synthetic products at the forefront.
- End-to-end CRO services, encompassing template production, in vitro transcription, capping and tailing, LNP encapsulation, and quality control.
- Comprehensive and top-notch mRNA quality inspection platforms, supported by methods such as capillary electrophoresis, high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), and more."
mRNA BsaI
mRNA BsaI
Bsa I , an IIs restriction endonuclease restriction endonuclease, is derived from a recombinant E.coli strain that carries the cloned and modified BsaI gene from Bacillus stearothermophilus. which have the same specificity as native enzymes and reduced star activity.
Its recognition sequence and cleavage sites are as follows:
5’······GGTCTC(N)···············3′
3’······CCAGAG(NNNNN)······5
BsaI
| Catalog | Size |
| HBP001306 | 1KU |
| HBP001307 | 5KU |
| HBP001308 | 200KU |
| HBP001309 | 2000KU |
Product component
| Component | 1KU | 5KU |
|---|---|---|
| BsaI（20 U/μL） | 50 μL | 250 μL |
| 10 x BsaI Buffer | 1.25 mL | 1.25 mL |
| Component | 200KU | 2000KU |
|---|---|---|
| BsaI （20 U/μL） | 10 mL | 100 mL |
| 10 x BsaI Buffer | 5×10 mL | 5×100 mL |
Storage
Store at -20 ℃, valid for 1 year（ Avoid repeated freeze-thaw cycles）
Storage buffer
10 mM Tris-HCl, 200 mM NaCl, 1 mM DTT 0.1 mM EDTA, 200 µg/ml Recombinant Albumin,50% Glycerol. (pH 7.4 @ 25°C).
Unit Definition
One unit is defined as the amount of enzyme required to digest 1 µg of Internal control DNA in 1 hour at 37°C in a total reaction volume of 50 µL.
Quality Control
Protein Purity Assay (SDS-PAGE): The purity of Bsa I was ≥ 95% determined by SDS-PAGE analysis using Coomassie Blue detection.
RNase: 20 U of Bsa I with 1.6 μg MS2 RNA for 16 hours at 37 ℃ yields no degradation as determined by agarose gel electrophoresis.
Non-Specific DNase Activity: 20 U of Bsa I with 1 μg PhiX174 DNA for 16 hours at 37 ℃ yields no excess DNA as determined by agarose gel electrophoresis.
Star activity: 40 U of Bsa I with 1 μg λ DNA/HindIII for 16 hours at 37 ℃ results in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.
Ligation and Recutting: After digestion of 1 μg λDNA/HindIII with 20 U BsaI, DNA fragments can be ligated with T4 DNA ligase at 16ºC. And these ligated fragments can be recut with BsaI.
E.coli DNA: 2 U of Vaccinia virus Capping Enzyme is screened for the presence of E. coli genomic DNA using TaqMan qPCR with primers specific for the E. coli 16S rRNA locus. The E. coli genomic DNA contamination is≤1 E.coli genome.
Bacterial Endotoxin: LAL-test, according to Chinese Pharmacopoeia IV 2020 edition, gel limit test method, the general rule (1143). Bacterial endotoxin content should be ≤10 EU/mg.
Reaction system and conditions
| Component | Volume |
|---|---|
| BsaI（20 U/μL） | 1 μL |
| DNA | 1 μg |
| 10 x BsaI Buffer | 5 μL |
| dd H2O | Up to 50 μL |
Incubate at 37°C for 15-30 minutes Heat Inactivation：80°C for 20 min
Product features
1.High activity
2. Fast digestion
3.Low star activity , ensuring accurate cutting like “scalpel”; Without BSA and animal-origin free.
Methylation Sensitivity
dam methylation: Not Sensitive;
dcm methylation: Impaired by Some Combinations of Overlapping;
CpG Methylation: Blocked by Some Combinations of Overlapping.
Notes on use
- The volume of enzyme ≤ 1 / 10 of the reaction volume.
- Star activity may occur when glycerol concentration is more than 5%.
- Cleavage activity may occur when Substrate below the recommended ratio.
mRNA BspQI
Description
BspQI can be recombinantly expressed in E. coli that can recognize specific sites and are produced under GMP standards. It is widely used in gene mapping, cloning and other fields. This enzyme rapidly cuts DNA for efficient gene linearization.
BspQI
| Cat. No. | Size |
| HBP001401 | 0.5 KU |
| HBP001402 | 2.5 KU |
| HBP001403 | 100 KU |
| HBP001404 | 1000 KU |
Product Component
| Component | HBP001401 | HBP001402 |
|---|---|---|
| BspQI (10 U/μL) | 50 μL | 250 μL |
| 10 x Buffer | 1.25 mL | 1.25 mL |
| Component | HBP001403 | HBP001404 |
|---|---|---|
| BspQI (10 U/μL) | 10 mL | 100 mL |
| 10 x Buffer | 12.5 mL | 125 mL |
Shipping and Storage conditions
The product should be shipped ≤ 0℃； Stored at -20℃ condition.
Unit Definition
One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 50°C in a total reaction volume of 50 µl.
Recognition Site
Quality Control
Purity≥95%, no DNase, RNase activity, Host DNA residue≤100pg/mg, Host protein residue ≤50ppm, Endotoxin residue≤10EU/mg, No protease activity, sterile, no mycoplasma.
Reaction system and conditions
| Component | Volume |
|---|---|
| BspQI(10 U/μL) | 1 μL |
| DNA | 1 μg |
| 10 × Buffer | 5 μL |
| dd H2O | Up to 50 μL |
Reaction time：50℃, 15~30 min。
Product Features
- The restriction endonuclease has high enzymatic activity and completes the digestion quickly.
- Low non-specific endonuclease activity to ensure precise “scalpel” cutting.
- No BSA system, reducing heat source pollution.
Application
- RNA Vaccine Preparation
- Restriction Enzyme Digestion
- Fast Cloning.
