Gentaur Ultra Nuclease

HMD5501

Product can degrade all forms of DNA and RNA and digest them into. a length of 3-5 base

SKU                                            Pack Size  

• G-CAS 9025-65-4 HMD5501-5KU  5000 UNITS     €335
• G-CAS 9025-65-4 HMD5501-25KU 25000 UNITS  €1335

ME  UltraNuclease

1. The ULTRA Nuclease Benzoase like Nuclease, versatile nuclease, is used as a broad spectrum nuclease.
2. Endonuclease non-limiting nucleic acid will cut and degrade DNA and RNA to remove biological products and nuclear acids.

The Gentaurs's production process of UltraNuclease is completed in the Gen9 GMP production facility with FDA quality inspection of the Pharmacopoeia.

The enzyme activity of protease-free, endotoxin free, and any viral contaminants.

Wha tis the use of Ultra Nuclease?

The use of Ultranucease or benzoase is removing residual nucleic acid.

How does GEN9 produces Ultra Nuclease?

We express UltraNuclease as nonspecific endonuclease in recombinant E. coli form from Serratia Marcescen and expressed.

What is the action of Gen9 Ultranuclease?

GEN9 Nucleases from Escherichia coli (E. coli) catalyze nonspecific endonucleolytic cleavage of DNA and  SARS-COV2 RNA.

Double-stranded, single-stranded, linear, circular, naturally occurring, or denatured DNA are processed to yield 5'-monophosphate oligonucleotides of 2-5 bp.

What conditions does the Gen9 Ultra Nuclease needs to Remove Covid RNA and DNA?

1. This can tak eplace is  followingconditions: 6 M Urea， 0.1 M Guanidine HCl，0.4% Triton X-100，0.1% SDS，1 mM EDTA，1 mM PMSF.
2. This product has been widely used for nucleic acid digestion for RT-PCR of Covid.

We expressed the recombinant nucease from Escherichia coli (E. coli) with Purity ≥ 95%.

How to reduce with Nuclease cell supernatant or cell lysate viscosity?

1. Increasing protein purification efficiency and enhancing protein functional research will prevent clumping of human peripheral blood mononuclear cells (PBMC) in cell therapy and vaccine development.
2. The product is provided in the form of a sterilized reagent, eluted in buffer（20 mM Tris-HCl pH 8.0，2 mM MgCl2，20 mM NaCl，50% glycerin）, with the appearance of colorless, transparent liquid.

How UltraNuclease compares to Benzonase Nuclease?

Both are ≥250 units/μL, ≥99% (SDS-PAGE), recombinant  E. coli in glycerol solution, ultrapure grade

Gen9 UltraNucease Synonym(s):

1. Endonuclease from Serratia marcescens
2. CAS Number: Gen 9025-65-4
3. Enzyme Commission number: 3.1.30.2 (BRENDA, IUBMB)
4. MDL number: MFCD00131010
5. NACRES: NA.54

Gen9 Ultra Nuclease Specifications

• Host: Escherichia coli (E. coli)
• Molecular Weight: 26.5 kDa
• Isoelectric point: 6.85
• Purity:≥ 95%（SDS-PAGE）
• Enzyme Activity: ≥ 250 U/μL
• Specific Activity:≥1.5×106 U/mg protein
• Optimum pH:8.0（working pH range: 6-10）
• Optimum Temperature: 37℃（working temperature range: 0-42℃）
• Cofactor: 1-10 mM Mg2+
• Protease: Undetected
• Storage Buffer: 20 mM Tris-HCl pH8.0，2 mM MgCl2，20 mM NaCl，50% glycerin
• Unit Definition: The definition of one Activity unit (U) is the amount of enzyme used to change the absorption value of △A260 by 1.0 in 30 minutes in a 2.625 mL reaction system at 37℃ with a pH of 8.0 (equivalent to complete digestion of 37 μg salmon sperm DNA into oligonucleotides).
• Storage and Transportation: Transport with ice packs. Storage at -20℃, Valid for 1 year. If the product is opened and has been stored at 4℃ for more than a week, we recommend filtering the product to prevent microorganism contamination.

Important Notes

Please wear PPE, such lab coat and gloves, to ensure your health and safety.

How do we compare to DENARASE?

Enzyme engineering and bioprocess development for DENARASE for
Use in pharmaceutical products.
DENARASE, a so-called endonuclease, is an enzyme that is also used to remove DNA in the purification of as well
Vaccines is used.

 Biologic License Applications (BLA), Investigational New Drugs (IND) andNew Drug Applications (NDA)

The FDA has accepted UltraNuclease Amount at its  final Concentration and purity SOP

Ultra Nuclease Production Protocol

1. Sample Collection

Adherent cells: remove the medium, wash the cells with PBS, and remove the supernatant.

Suspension cells: collect the cells by centrifugation, wash the cells with PBS, centrifuge at 6,000 rpm for 10 min, and collect the pellet.

Escherichia coli: collect the bacteria by centrifugation, wash once with PBS, centrifuge at 8,000 rpm for 5 min, and collect the pellet.

2. Sample Treatment

Treat the collected cell pellets with lysis buffer at the ratio of mass (g) to volume (mL) 1: (10-20), or by mechanical or chemical methods on ice or at room temperature (1 g of cell pellet contains about 109 cells).

3. Enzyme Treatment

Add UltraNuclease according to the ratio of 250 Units to digest 1 g of cell pellets. Please refer to the “Recommended Treatment Time” form above to choose the duration of the treatment.

4. Supernatant Collection

Centrifuge at 12,000 rpm for 30 minutes and collect the supernatant.

Note: If the solution is acidic or alkaline, or contains high concentrations of salt, detergents, or denaturants, please increase the enzyme dosage or extend the treatment time accordingly.

Almighty Nuclease for broad application RNA removal 

1. Reduce the viscosity of nucleic acid released during cell/bacterial lysis, and it is suitable for any lysis method.
2. In large-scale chromatographic purification, avoid reducing the effective load of protein due to the adsorption of a large amount of nucleic acid on the chromatographic medium, thereby reducing the purification yield.
3. In ELISA, two-dimensional electrophoresis and western blot analysis, improve the resolution and recovery rate.
4. Elimination of exogenous nucleic acid residues in biological products such as recombinant proteins and vaccines.

Gen9 Ultranuclease Product Features

The specific activity of the nuclease enzyme is ≥ . 1 .5×10 . 6 the U- / mg (universal substrates absolute quantification)

Enzymatic activity:  The enzyme used to change the absorption value of △ A 260 by 1.0 (equivalent to complete digestion of 37 μg salmon sperm DNA into oligonucleotides) within 30 minutes in a 2.625 mL reaction system under 37°C , pH 8.0 reaction conditions The amount is defined as an activity unit ( U )

Nuclease Enzyme purity ≥99%(HPLC)

The influence of different reaction conditions on enzyme activity

Operating conditions for UltraN uclease

Storage and freeze-thaw stability

When stored at -20 ℃ for 8 weeks, there was no significant change in enzyme activity; when stored at 4 ℃ and 37 ℃ for 8 weeks, the enzyme activity decreased by about 20% and 25%, respectively .

-80℃~25℃ repeated freezing and thawing 10 times, the enzyme activity will not be affected .

Enzyme activity

250-300 U /u L

Universal substrate method

Than live

≥1.5×10 6 U/mg

Universal substrate method

Protein purity

≥99%

SDS-PAGE

Protease

Meet the requirements

Universal substrate method

1. Bacterial endotoxin
2. Meet the requirements
3. Gel method
4. Host protein
5. ELISA method
6. Host DNA
7. qPCR method
8. Sterility testing
9. Aseptic growth
10. General Principle 1101
11. Pathogen detection
12. Negative (not detectable)
13. Refer to internal method
14. Mycoplasma detection
15. Negative (not detectable)
16. Refer to internal method
17. Heavy metal detection
18. Meet the requirements
19. General Principle 0 821

UltraNuclease E LISA kit ( Almighty Nuclease Detection E LISA k it)

In normal purification steps, the totipotent nuclease is easily removed as an impurity. In order to detect the residue of totipotent nuclease, Yisheng has developed the supporting UltraNuclease ELISA kit , which can accurately detect the residue of totipotent nuclease in the sample. The kit has excellent performance in linearity, repeatability, recovery rate and specificity .

 linear range:0.047 ~. 3 ng / mL,R & lt2>0.99

Compare to Benzo Nuclease

1. Description:
2. Benzo Nuclease is a kinds of no-specific endonuclease. The Benzo Nuclease genetically engineered endonuclease from Serratia marcescens.
3. Benzo Nuclease degrades all kinds of DNA and RNA (double stranded, single stranded, linear and circular and supercoil) but without proteolytic activity. It is effective over a wide range of conditions and possesses an exceptionally high specific activity.
4. Benzo Nuclease completely digests nucleic acids to 5'monophosphate terminated oligonucleotides 2 to 5 bases in length (below the hybridization limit), which is ideal for removal of nucleic acids from recombinant proteins, enabling compliance with FDA guidelines for nucleic acid contamination. The ability of Benzo Nuclease to rapidly hydrolyze nucleic acids makes the enzyme an excellent choice for viscosity reduction to reduce processing time and increase yields of protein. For example, the enzyme is compatible with BugBuster® and PopCulture® Protein Extraction Reagents and can therefore be added along with these reagents to eliminate viscosity and remove nucleic acids from E. coli extracts. The enzyme consists of two subunits of 30 kDa each. It is functional between pH 6 and 10 and from 0-42°C and requires 1-2mM Mg2+ for activation. The enzyme is also active in the presence of ionic and non-ionic detergents, reducing agents, PMSF (1 mM), EDTA (1 mM) and urea (relative activity depends on specific conditions). Activity is inhibited by>150 mM monovalent cations,>100 mM phosphate,>100 mM ammonium sulfate, or>100 mM guanidine HCl.
5. Source: Serratia marcescens
6. Molecular weight: 27. 9 KD (SDS)
7. Purity: ≥90%, (Ultra≥99%)(SDS-PAGE)
8. PI: 6. 85
9. Optimum pH: 8. 0
10. Optimum temperature: 37℃
11. Cofactor: 1～10mM Mg2+
12. Form: Liquid enzyme is the clear Solution(Contain Glycerin)
13. Solid enzyme is the white lyophilized powder.
14. Concentration: 1000 U/µl.
15. Activity: ≥1000U/μl
16. Specific activity: ≥1. 0x106U/mg protien
17. Protease: Not detectable
18. Microbial limit: aerobe<10CFU/100 KU Yeast and mold: <10CFU/100 KU
19. Endotoxin test: (Gel Clot LAL Assay)<0. 25EU/1000U
20. Storage Buffer:
21. 20 mM Tris-Cl(pH 8. 0), 2 mM MgCl2, 2 mM NaCl, 50%Glycerin
22. Dilution Buffer:
23. 20 mM Tris-Cl(pH 8. 0), 2 mM MgCl2, 2 mM NaCl.
24. Unit definition:
25. One unit is defined as the amount of enzyme that causes a ΔA260 of 1. 0 in 30 minutes, which corresponds to complete digestion of 37 µg DNA.
26. Applification:
27. Its high intrinsic activity and broad substrate tolerance make the endonuclease an ideal tool in a variety of biotechnological and pharmaceutical applications:
28. removal of nucleic acid from protein samples
29. Elimination of nucleic acids from recombinant proteins
30. Purification of protein fragments from inclusion bodies;
31. Sample preparation in western blotting or two-dimensional gel electrophoresis
32. Viscosity reduction in protein extracts.
33. Shipping and Handling:
34. Liquid enzyme: Ship with Blue Ice, Storage at -20℃
35. lyophilized powder: Ship at room temperature, storage blow 4℃

Is Benzoase and UltraNuclease GMP-grade?

Both are.

1. UltraNuclease GMP-grade Almighty Nuclease
2. Product number: 20157ES25
3. Product Specifications: 25 KU

Other Specifications

• Storage and transportation: ice bag transportation. Store at -20℃, valid for 1 year. If it is left at 4℃ for more than a week after opening the package, it is recommended to filter and sterilize to prevent microbial growthIn stock status: In stockRange of use: for scientific research use only, not for clinical useClick here to buyOnline QQ: I am 2694086042, what can I do for you?Biological Online Statement: The information displayed above is provided by the company itself, and the authenticity, accuracy and legality of the content are the responsibility of the publishing company. Bio does not undertake any guarantee responsibility.

Detailed Description

The UCF . The ME ® the U- LTRA N uclease the GMP -g Rade versatile nuclease

pUltran uclease the GMP-Grade versatile nuclease

Product Description

UltraN uclease ( omnipotent nuclease ) , also known as non-restrictive endonuclease, broad-spectrum nuclease ; is a non-specific endonuclease derived from Serratia Marcescen , which can cut between any nucleotides in the chain, The nucleic acid is completely digested into 5 -monophosphate oligonucleotides with a length of 2-5 bases , which can be used under a very wide range of conditions ( 6 M Urea , 0.1 M Guanidine HCl , 0.4% Triton X - 100 , 0.1 % SDS , 1 mM EDTA , 1 mM PMSF ) degrade various forms of (double-stranded, single-stranded, linear, circular, natural or denatured) DNA and RNA , and are widely used to remove nucleic acids in biological products.

The product in the genetically engineered Escherichia coli (E. coli) are expressed and purified GMP preparation environment, not only in scientific research to reduce cell supernatants and cell lysates of viscosity , protein purification improve efficiency and functional studies; further can be used in virus purification, vaccine production , protein and polysaccharide in the pharmaceutical industry as a nucleic acid-removing residual host agent, the host residual nucleic acids down to picograms ( PG ) level of biological products to improve the efficacy and safety of; cell therapy can be effectively prevented and vaccine Research and human peripheral blood mononuclear cells ( P the BMC ) clumping.

This product is provided as a sterile liquid enzyme stored in the buffer ( 20 is mM Tris-HCI the pH 8.0 , 2 mM MgCI2 2 , 2 0 mM NaCI , 50% glycerol), colorless transparent liquid.

Transportation and storage methods

Ice pack transportation. Store at -20℃ , valid for 1 year. If the package is opened and placed at 4°C for more than a week, it is recommended to filter and sterilize to prevent microbial contamination.

Precautions

For your safety and health, please wear lab coats and disposable gloves for operation.

Recommended use conditions

Condition parameter

Optimal conditions

Effective conditions

Mg 2+

1-5 mM

Instructions

1. Sample preparation

Adherent cells: the medium was removed with PBS clean washed cells, the supernatant was removed.

Suspension cells: collect cells by centrifugation, wash the cells with P BS , centrifuge at 6,000 rpm for 10 min , and collect the pellet.

E. coli: bacterial cells were collected by centrifugation, with P the BS washed 1 times, 8,000 RPM centrifuge . 5 min , the precipitate was collected.

2. Sample processing

The collected cell pellets are lysed according to the ratio of mass ( g ) to volume ( mL ) 1 : (10~20) . Cells can also be lysed by mechanical or chemical methods on ice or at room temperature ( 1 g of cells is about It is 10 . 9 th) .

3. Adding enzymes

1 ) adding an appropriate amount M GCL 2 The reaction system of Mg 2+ concentration was adjusted at 1 -5 mM within range, P H was adjusted to 8-9 .

2 ) according to 250 Units digested 1 g ratio to add the cell pellet Ultran uclease , . 3 . 7 deg.] C incubation . 3 0 min or more. You can also choose the addition plan according to the recommended dosage in the above table, increase the amount of enzyme within a certain range, and reduce the time required for digestion.

[ Note ] versatile nuclease activity by the ion concentration, reaction temperature and P H Effects of other factors, the optimum concentration of initial exploration recommended use.

4. Obtain the supernatant

Centrifuge at 12,000 rpm for 30 minutes to obtain the cell lysate supernatant, and then perform subsequent related experiments.

[ Note ] If the solution is a high salt solution, acidic or alkaline, detergent containing a high concentration of denaturant, the enzyme should be increased by the amount or extended incubation time.

Gen9 Biological Online Statement:

The information displayed above is provided by the company itself, and the authenticity, accuracy and legality of the content are the responsibility of the publishing company. Bio does not undertake any guarantee responsibility.

Good repeatability between batches

In the sample is added a fixed concentration of Ultra Nuclease , with . 3 different batches of E LISA kit for detecting, in a nucleic acid enzyme concentration 3 ~ 0.186 ng / mL , between the detection recoveries were in the 70% to 110% between , C V < . 1 0% .

Recovery rate of simulated sample

For different concentrations of Ultra Nuclease nuclease samples, the recovery rate is between 70% and 130% , which is better than other brands.

Strong specificity

We evaluated the non-specific binding of the pluripotent nuclease antibody to 8 common samples. The results of the interference group and the control group overlapped, and no interference was seen.

UltraNuclease addition

It depends on the sample you are dealing with. When the virus is purified, it is usually added after the virus is clarified; when the E. coli protein is purified, it can be added when the bacteria is lysed; when the problem of cell clumping is handled, it can be directly added to the culture medium to co-culture with the cells.

Under reaction temperature cannot reach 37℃, how to use it?

The enzyme activity of UltraNuclease is affected by temperature, processing time, and enzyme activity unit. If the temperature cannot reach 37°C, the amount of enzyme can be appropriately added, or the incubation time can be extended.

Is UltraNuclease compatible with protease inhibitors?

It is compatible, but it should be noted that many protease inhibitors contain EDTA. When EDTA starts to be higher than 1mM, EDTA will inhibit part of the nuclease activity.

How to remove UltraNuclease?

It can be removed by TFF, dialysis and chromatography column.

Can UltraNuclease ELISA kit be used for the detection of residues of other brands of Almighty Nuclease?

We do not recommend this. ELISA kit is completely developed based on UltraNuclease, which can accurately quantify UltraNuclease. Other brands of Almighty Nuclease may be different from UltraNuclease in terms of sequence and production process, and the test results may be inaccurate.

UltraNuclease GMP-grade Totipotent nuclease

UltraNuclease ELISA kit