1EzNUCLEASE enzyme

• Easier to handle and purify cell lysate
• Highly active and purified
• No detectable protease activity

1EzNUCLEASE DNA & RNA Endonuclease is a proprietary E. coli preparation of the recombinant Serratia macescens extracellular endonuclease. This highly active enzyme digests single and double stranded nucleic acids (DNAs and RNAs) into 5'-phosphorylated oligonucleotides that are 1-4 bases long. The 1EzNUCLEASE is a 27 kDa highly purified homodimer (>99%), is highly active, and has no detectable protease activity. The 1EzNUCLEASE endonuclease makes it easier to handle and purify cell lysate components because it dramatically reduces sample viscosity via nucleic acids digestion.

50 mg of salmon sperm DNA was incubated with the indicated units of 1EzNUCLEASE and a commercially available brand of nuclease at 37oC for 30 min in a buffer of 50 mM Tris-HCl, pH 8.0 and 1 mM MgCl2. DNA digestion was monitored by agarose gel.

Shipping Storage
Shipped on blue ice
Store at -20ºC; stable for 6 months

MATERIALS AND METHODS A-Specifications - Activity: One unit of 1EzNUCLEASE converts 1.0 OD260 of salmon sperm DNA into acid-soluble nucleotides in 30 minutes at 37ºC. - Specific Activity: >1.3 x 106 units/mg (equivalent to >3 x 106 Kunitz units/mg, 100-fold more active than the purified bovine Dnase I). - Reaction buffer: 50 mM Tris-HCl, pH 8.0, 1mM MgCl2. - Storage buffer: 50 mM Tris-HCl, pH 8.0, 50mM NaCl, 5mM MgCl2, 50% glycerol. - Stability: Stable in storage buffer at 37ºC for one week without any loss of activity. Can be stored at -20ºC or flash frozen and stored in liquid nitrogen or -70ºC. B- Digestion Conditions 1. To reduce viscosity of cell lysates, start with 10-100 units of 1EzNUCLEASE per gram of cell paste. 2. The efficiency of digestions and reduction in viscocity varies with buffers, cell types, and cell lysis methods used. If needed, increase the amount of 1EzNUCLEASE to 1,000 units per gram of cell paste. NOTE: the total amount of 1EzNUCLEASE present in 1,000 units is small enough (<1μg/ml) not to cause any downstream complications.