UltraNuclease, also known as non restrictive endonuclease or broad-spectrum nuclease; It is a non-specific endonuclease derived from Serratia Marcesen that can cleave between any nucleotide in the chain, completely digesting the nucleic acid into 2-5 base length 5 '- monophosphate oligonucleotides. It can degrade various forms of DNA and RNA under a wide range of conditions and is widely used to remove nucleic acids from biological products.
【Properties】
1、This product has been extensively fermented, expressed, and purified using Escherichia coli (E. coli), with a purity of over 95%. It can be used in scientific research to reduce the viscosity of cell supernatants and lysis liquids, improve protein purification efficiency and functional research, and effectively prevent the aggregation of human peripheral blood mononuclear cell (PBMC) in cell therapy and
vaccine research.
2、This product is provided in the form of sterile liquid enzyme and stored in buffer solution (20 mM Tris HCl pH 8.0, 2mM MgCl2, 20mM NaCl, 50% glycerol), a colorless and transparent liquid.
【Stability】
UltraNuclease can maintain high stability and digestive activity of nucleic acids under a wide range of conditions, making them compatible with various cell lysis liquids, such as RIPA, or protein extraction reagents containing multiple ionic and non-ionic detergents and reducing agents. For example, guanidine hydrochloride with a concentration of>100mM will completely inhibit activity, 1mM EDTA will partially inhibit activity, 5mM EDTA will cause a 90% loss of activity, and 1mM PMSF will not inhibit nuclease activity.
【Usage】
1、Cell processing:
a. Remove the culture medium from adherent cells, wash with PBS, and add 1-2 μL UltraNuclease to 1mL of RIPA lysis solution (or other mammalian cell lysis solution) at room temperature or ice for 5-30 minutes, collect the lysis liquid, centrifuge and obtain the supernatant for downstream experiments.
b.After collecting suspended cells by centrifugation, add 1mL of RIPA lysis fluid (or other mammalian cell lysis fluid) and 1-2 μL UltraNuclease to the centrifuge tube at room temperature or ice for 5-30 minutes, collect the lysate, centrifuge, and obtain the supernatant for downstream experiments.
2、Organizational samples： After grinding 30-100 mg of animal or plant tissue thoroughly, add 100-200 μL lysis fluid, while adding 0.5-1 μL UltraNuclease at room temperature or ice for 5-30 minutes, collect the lysis liquid, centrifuge and obtain the supernatant for downstream experiments.
3、Escherichia coli or other bacteria：After bacterial centrifugation collection, they are lysed or ground to pieces with lysis liquid, and 0.5-1μL UltraNuclease is added to every 1 mL, incubate at room temperature for 30 minutes, collect the lysis liquid, centrifuge and take the supernatant for downstream experiments.
【Application】
1、 Remove exogenous nucleic acids from the production process of vaccine products to improve their biological safety;
2、Removing the nucleic acid entangled on the surface of virus particles is beneficial for the release and purification of virus particles;
3、 Degradation of nucleic acids in cell lysis fluid, reduction of liquid viscosity, and increase of recombinant protein production;
4、Process samples used for ELISA, electrophoresis, or column chromatography purification to improve resolution and recovery.
【Attention】
For your safety and health, please wear Lab coat and disposable gloves.
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