| Source | Isolated from Proteus vulgaris 84. |
| Description | Endonuclease I hydrolyzes double- and single-stranded nucleic acids to oligonucleotides of 3-5 nucleotide lenghts with 5` - terminal phosphates.
Application: DNA and RNA degradation
|
| Unit definition | One unit is defined as the amount of enzyme required to hydrolyze 1 μg of Lambda DNA in a total reaction volume of 50 μl in 30 minutes at 37°C.
Unit Assay Conditions: 20 mM Glicin-NaOH (pH 9.5), 100 mM NaCl, 25 mM MgCl2, 1 mM 2-mercaptoetanol
|
| Reaction buffer | SE-buffer Endonuclease I, (20 mM Glycine-NaOH (pH 9.5 at 25°C); 25 mM MgCl2; 100mM NaCl; 1mM 2-mercaptoethanol) |
| Optimal temperature | 37oC |
| Storage conditions | 10 mM Tris-HCl (pH 7.4); 250 mM NaCl; 0,2 mM EDTA; 100 μg/ml BSA; 7 mM 2-mercaptoetanol ; 50% glycerol. Store at -20°C. |
| Quality control | 10 units of Endonuclease I is incubated with 20 ng of a single or double-stranded synthetic
oligonucleotide in a 10 μl reaction mixture with the SE Endonuclease I Buffer for 3 hours at 37°C.
Phosphatase contamination is determined by the loss of radioactivity from original 5` 32P-labeled oligonucleotide or products of 32P-labeled oligonucleotide digestion with Endonucleases I as determined by denaturating polyacrylamide gel electrophresis and subsequent autoradiography. |
| MSDS: | Download MSDS as PDF |
Runs:
1. double-stranded synthetic oligonucleotide 5`p*GTGCATGGCGCCTATGCG3`/ 3`CACGTACCGCGGATACGC5`
2. double-stranded synthetic oligonucleotide 5`p*GTGCATGGCGCCTATGCG3`/ 3`CACGTACCGCGGATACGC5` plus Endonuclease I
3. single-stranded synthetic oligonucleotide 5`p*GTGCATGGCGCCTATGCG3`plus Endonuclease I
4. double-stranded synthetic oligonucleotide 5`p*GTGCATGGCGCCTATGCG3`/ 3`CACGTACCGCGGATACGC5` plus endonucleases Fai I (recognition sequence YA^TR)
5. single-stranded synthetic oligonucleotide 5`p*GTGCATGGCGCCTATGCG3`
6. single-stranded synthetic oligonucleotide 5`p*GTGCATGGCGCCTATGCG3`plus Alkaline Phosphatase
labeled strand is marked by a symbol <*>
|  |