Activation of DNA primer for DNA polymerase as an ultrasensitive assay for a subgroup of enzymes with deoxyribonuclease activity
| Autor: | Gary J. Fischman, George P. Studzinski |
|---|---|
| Rok vydání: | 1974 |
| Předmět: |
Swine
DNA polymerase DNA polymerase II Biophysics Thymus Gland Nucleic Acid Denaturation Tritium Biochemistry Chromatography DEAE-Cellulose Micrococcus chemistry.chemical_compound Escherichia coli Methods Animals Pancreas Molecular Biology Polymerase Cell Nucleus Deoxyribonucleases DNA clamp biology Phosphoric Diester Hydrolases Venoms Microchemistry Snakes DNA Cell Biology Deoxyribonuclease activity Molecular biology chemistry Evaluation Studies as Topic Sea Urchins DNA Nucleotidyltransferases biology.protein Cattle DNA polymerase I Primer (molecular biology) HeLa Cells |
| Zdroj: | Analytical Biochemistry. 58:449-458 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/0003-2697(74)90213-9 |
| Popis: | Pretreatment of native DNA with nucleases which hydrolyze phosphodiester bonds with liberation of 3′-hydroxyl terminal groups is known to increase the rate of incorporation of nucleotides into DNA by E. coli DNA polymerase I and similar enzymes. Concentration ranges and conditions for this reaction have been established which allow specific detection and quantitative assay of such nucleases. The method permits detection of as little as 0.1 pg of pancreatic deoxyribonuclease, is simple, and requires only commercially available components. It has been utilized to monitor purification of a nuclease from HeLa cell nuclei at concentrations which could not be detected by other methods. |
| Databáze: | OpenAIRE |
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