Pannarase Nuclease is an endonuclease genetically engineered from Serratia marcescens, and it is able to degrade various types of DNA and RNA into 5'-monophosphate oligonucleotides of 3 to 5 base lengths. It is also known as "Nuclease". It is widely used in viral purification processes for the development of vaccines and cell and gene therapies.
Properties:
Item | Description |
Specification | 100,000/500,000/5,000,000 U/tube |
Molecular weight | 27kDa |
Purity | ≥99% (SEC-HPLC) |
Specific activity | ≥2x106 U/mg |
Optimal pH | 8.0 |
Optimal temperature | 37°C |
Cofactor | 1-10 mM Mg2+ |
Buffer | 20 mM Tris-HCl pH 8.0, 2 mM NaCl, 2 mM MgCl2, 50% (v/v) glycerol |
Storage temperature | -20°C, do not freeze and thaw repeatedly (It does not freeze at -20°C due to the presence of glycerin) |
Definition of active unit: when cleaving herring sperm DNA as the substrate (herring sperm DNA, Sigma, catalog number D7290) at 37°C, pH 8.0, one active unit of Pannarase Nuclease will produce a decrease of 1.0 in A260 (absorbance value at 260 nm) per 30 minutes (equivalent to a complete cleavage of 37 μg DNA).
Features:
SEC-HPLC：（purity 99.72%）
Size exclusion chromatography showed high purity with no aggregates produced
LC-MS test：
LC-MS analysis indicates that the protein molecular weight is consistent with the theoretical value, featuring no impurity peaks, showing homogeneous structure
Application case study:
Enzyme digestion activity on plasmid DNA
Comparison of enzymatic digestion effect of different brands of nucleases
(Reaction condition: reaction system of 20 uL with plasmid DNA concentration of 1 mg/mL)
Transportation and Storage:
Transport on dry ice. When stored at -15℃ ~ -25℃, the validity period is 2 year.
Recommended usage conditions:
Reaction condition | Optimal conditions* | Effective condition** |
Mg2+ | 1-2 mM | 1-10 mM |
pH | 8.0-9.2 | 6.0-10.0 |
Na+/K+ concentration | 0-20 mM | 0-150 mM |
Temperature | 37℃ | 4-42℃ |
(Dithiothreitol)DTT | 0-100 mM | 0-300 mM |
2-mercaptoethanol | 0-100 mM | 0-300 mM |
PO43- | 0-10 mM | 0-100 mM |
*“Optimal condition” means the condition under which Pannarase nuclease maintains >90% activity
**“Effective condition” means the condition under which Pannarase nuclease maintains >15% activity
Usage:
1.For fragmentized liquid of E. coli
For the purpose of viscosity reduction, the amount of nuclease to be added must be decided according to the concentration of bacteria in the fragmentized liquid. For the concentration of bacteria at 50%, the recommended amount of nuclease to be added is 1:1000-5:1000, i.e. 500,000-2.5 million U/L. For the concentration of bacteria at 5%, addition can be made in the ratio of 1:10,000 to 5:10,000.
2.For the cell lysate
Addition of 500 units of nuclease can be made for every 106-107 cells.
3.For purified adenovirus or viral vaccine
Due to the relatively small amount of DNA, it can be added at a final concentration of 0,01% to 0,05%, that is, 5 units per mL.
Attention:
1.This enzyme works best at pH 8.0-11.0, 37°C, for 30 min.
2. Magnesium ion is necessary for enzyme activity and the recommended concentration is 2 mM. If there is a metal chelator such as DETA in the sample, magnesium ion should be accordingly removed or added.
3.Due to the stable activity, it is appropriate to place the enzyme at room temperature for a short period of time, but it should be placed in the refrigerator at -20°C for long-term storage. The enzyme should be used as soon as possible after dilution with glycerol-free buffer, and it is not recommended to use it after frozen storage.
4. For the sole purpose of viscosity reduction, nuclease can be added directly and stirred slowly for 30 min at room temperature.
Order Imformations: Cat.NO. Name Grade Size CY002F0010 Pannarase Nucleases RUO 100 kU CY002F0050 Pannarase Nucleases RUO 500 kU CYG002F0010 Pannarase Nucleases GMP 100 kU CYG002F0050 Pannarase Nucleases GMP 500 kU CYG002F0500 Pannarase Nucleases GMP 5000 kU
Q:What is Pannarase Universal Nuclease
A:Pannarase is a non-specific nucleic acid endonuclease derived from Serratia marcescens, which can degrade any form of DNA and RNA (linear, circular, super spiral) into 3-5 bp oligonucleotides and maintain efficiency under a wide range of operating conditions
Q: What factors will affect the digestion effect of Pannarase omnipotent nuclease
A:The amount of enzyme added, reaction conditions (such as time, temperature, pH value), buffer environment (such as the presence of enzyme inhibitors), etc. The optimal usage of different samples requires experimental exploration
Q:What conditions will inhibit the activity of Pannarase omnipotent nuclease?
A:Pannarase omnipotent nuclease can maintain its activity under a wide range of conditions, and divalent cations (Mg2+) are crucial for its activity. When in a system containing the following substances, their activity is inhibited:>300 mM monovalent cations,>100 mM phosphates,>100 mM guanidine hydrochloride,>100 mM ammonium sulfate, etc.
When the system contains high concentrations of monovalent cations, it is recommended to choose Pannarase salt active nuclease.
Q:How to remove Pannarase omnipotent nuclease?
A:Conventional chromatography methods such as affinity chromatography and ion exchange chromatography can easily remove Pannarase universal nucleases. Other convenient purification methods, such as tangential flow filtration (TFF), are also suitable for removing Pannarase universal nucleases from biological products.
