Pannucleaseâ—‹R (EC:188.8.131.52) is a endonuclease that is produced and purified from E. coli. It has 245 amino acid residues with apparent molecular weight of ~29 kDa on a reducing SDS-PAGE gel. This endonuclease cleaves all single or double stranded DNA and RNA (either linear or circular) and is effective over a wide range of buffer conditions. It completely digests nucleic acids to 2–5 bases in length. It’s activity requires 1-2mM of Mg2+ and maintain high activity at temperature ranging 0ËšC to 42 ËšC.
Purity: ≥99% by Coomassie Blue staining method (Fig 1)
Specific activity: ≥1×106/mg orï¼ˆ1000U/ulï¼‰(Fig 2)
Endotoxin level: ≤0.02 EU/ug protein by LAL method.
Unit definition: Complete digestion of 36ug of DNA/RNA in a buffer of 50 mM Tris-HCl, pH 8.0, 1 mM MgCl2, at 37°C within 30 minutes
Storage (DO Not Store at -70 ËšC):
Liquid form: One month at room temperature (25 ËšC) or two years in –20 ËšC.
Lyophilized form: One year at room temperature (25 ËšC). Once it is reconstituted with water, store at -20ËšC for up to 2 years.
Example of application:
Removal of DNA/RNA contamination from product.
A typical bio-reactor generates large amount of cell mass but also creates DNA/RNA contamination due to cell death. In many cases it is desired to remove these contaminations before downstream processing. The range of DNA/RNA amount of contamination is usually from 0.5 to 50 ug/ml in the conditioned media or cells (2×107). We recommend for upstream treatment, add Pannucease at 1000 to 5,000 units per liter of culture to efficiently eliminate these DNA/RNA contamination (Fig 3). For downstream processing, significant lower amount of Pannuclease (ie. 100 to 1000 unit per liter) can be used.
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