Product info: Endonuclease I


Name
Endonuclease I
Cat. #E323E324
Package, u.a.400020000
Concentration, u.a./ml10000-5000010000-50000

SourceIsolated from Proteus vulgaris 84.
DescriptionEndonuclease 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 definitionOne unit is defined as the amount of enzyme required to hydrolyze 1 g of Lambda DNA in a total reaction volume of 50 l in 30 minutes at 37C.
Unit Assay Conditions: 20 mM Glicin-NaOH (pH 9.5), 100 mM NaCl, 25 mM MgCl2, 1 mM 2-mercaptoetanol
Reaction bufferSE-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 conditions10 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 -20C.
Quality control10 units of Endonuclease I is incubated with 20 ng of a single or double-stranded synthetic oligonucleotide in a 10 l 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 <*>