| Catalog # | R1992-03 |
| Source | Avian myeloblastosis virus particles |
| Storage Buffer | 200mM potassium phosphate (pH 7.2), 0.2% Triton X-100, 2mM DTT and 50% glycerol |
| Reaction Buffer (5X) | 250mM Tris-HCl (pH 8.3 @ 25°C), 250mM KCl, 50mM MgCl2, 2.5mM spermidine and 50mM DTT. |
| Unit Definition | One unit is the amount of enzyme required to catalyze the transfer of 1nmol of deoxynucleotide into acid-precipitable material in 10 minutes at 37°C. Reaction conditions are: 50mM Tris-HCl (pH 8.3), 8.75mM MgCl2, 40mM KCl, 10mM DTT, 0.1mg/ml BSA, 1mM radiolabeled dTTP and 0.25mM poly(A)400 and 0.25mM oligo(dT)50. |
| Quality Control | First-Stand cDNA Synthesis: First strand cDNAs, of 1.2 kb Control RNA is synthesized using 30 units of enzyme, 1ug of each template, an oligo(dT) primer and a radiolabeled dNTP. The minimum specification is the production of 120ng of first-strand cDNA. Full-length cDNA must be observed by gel electrophoresis and autoradiography. |
| Endonuclease Activity | 1ug of Type I supercoiled plasmid DNA is incubated with 25 units of enzyme in 50 mM Tris (pH 8.3), 40 mM KCl, 7mM MgCl2, 10 mM DTT for one hour at 37°C. Following incubation, the supercoiled DNA is visualized on an ethidium bromide-stained agarose gel to verify the absence of visible nicking or cutting. |
| Nuclease Activity | 50ng of radiolabeled DNA or RNA is incubated with 25 units of enzyme in 4mM Tris (pH 8.3), 3.2mM KCl, 0.56mM MgCl2, 0.8mM DTT for one hour at 37°C, and the release of radiolabeled nucleotides is monitored by scintillation counting of TCA-soluble material. Passing specifications is < 1% release for DNase and < 3% release for RNase. |
| Storage and Stability | May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for at least 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. |
| Source | Avian myeloblastosis virus particles |
| Purity | > 80% as judged by SDS-polyacrylamide gels with Coomassie blue staining. |
| Concentration | ~10u/ul |
|
| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
