Nuclease P1 (Nuclease 5'-Nucleotidehydrolase, 3'-Phosphohydrolase, NP1)

Blue Ice

4250

Supplied as a powder lyophilized from a solution containing 30mM sodium acetate buffer, pH 5.3 with 5mM ZnCl2 and 50mM sodium chloride. Enzyme suspension obtained after reconstitution.

Nuclease P1 (Nuclease 5'-Nucleotidehydrolase, 3'-Phosphohydrolase) from Penicillium citrinum, a zinc dependent glyco-enzyme consisting of 270 amino acid residues, hydrolyzes both 3'-5'-phosphodiester bonds in RNA and heat denatured DNA and 3'-phosphomonoester bonds in mono- and oligonucleotides terminated by 3'-phosphate without base specificity. Nuclease P1 is capable of hydrolyzing single stranded DNA and RNA completely to the level of mononucleoside 5’-monophosphates. The enzyme does not attack double-stranded nucleic acids, especially in the presence of more than 400mM of sodium chloride at pH 6.0.

Activity:
≥300U/mg protein using RNA substrate

Unit Definition:
One unit of nuclease activity is defined as the amount of enzyme that produces 1umole of acid soluble nucleotides from RNA based on E = 10, 600 for RNA hydrolysates per minute at 37°C, pH 5.3. One unit of the enzyme liberates 1umole of orthophosphate from 3’-AMP per minute at 37°C, pH 7.2

Optimum Temperature:
~70°C [< 60°C for long incubations]

Stable pH:
5.0-8.0

Reconstitution:
For maximum recovery of product, briefly centrifuge the original vial prior to opening. This will dislodge any enzyme that has adhered to the side or in the cap due to shipping. Reconstitute with sterile ddH2O. Allow the nuclease to rest on ice for ~10-15 minutes. Vortex gently to ensure there is uniform suspension (Nuclease tends to become layered towards the bottom of the vial). Just before addition to the reaction mixture, gently vortex the suspension.

Storage and Stability:
Lyophilized powder may be stored at -20°C. Stable for 12 months. Reconstituted product should be aliquoted and stored at -20°C. Reconstituted product is stable for 2-3 weeks at 4°C and 3-6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.