| 5'-G G T C T C(N)1^-3' |
|
| 3'-C C A G A G(N)5^-5' |
|
| Catalog # | E0373-55 |
| Concentration | 10u/ul |
| Source | E. coli that carries the cloned eco31IR gene from E. coli RFL31 |
| Reaction Buffer (10X) (E0373-55A) | Tris-HCl (pH 7.5), 10mM MgCl2, 50mM NaCl and 0.1mg/ml BSA. Incubate at 37°C |
| Restriction Enzyme Buffer A, 10X | 33mM Tris-acetate (pH 7.9), 10mM magnesium acetate, 66mM potassium acetate and 0.1mg/ml BSA. Incubate at 37°C. Supplied to simplify buffer selection for double digests. |
| Diluent Buffer | 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol, or Storage Buffer. |
| Storage Buffer | 10mM Tris-HCl (pH 7.5 at 25°C), 200mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol. |
| Overdigestion Assay | |
| No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with Eco31I (see Star Activity). |
|
| Ligation/Recutting Assay | |
| After 50-fold overdigestion (3u/ug DNA x 17 hours) with Eco31I, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.05uM. More than 95% of these can be recut. |
|
| Labeled Oligonucleotide (LO)Assay | |
| No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of restriction endonuclease for 4 hours. |
|
| Star Activity | |
| A large excess of enzyme (10u/ug DNA x 16 hours) may result in star activity. |
|
| Methylation Effects | |
| Sequence GAGACm5C is cleaved by Eco31I at least 50-fold slower than an unmethylated site. Overlapping Dcm or CG methylation may influence DNA cleavage. |
|
| Stability during Prolonged Incubation | |
| A minimum of 0.3 units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. |
|
| Thermal Inactivation | |
| Enzyme is inactivated by incubation at 65°C for 20min. |
|
| Digestion of Agarose-embedded DNA | |
| A minimum of 5 units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. |
|
| Number of Recognition Sites in DNA | |
| Lambda | 2 |
| PhiX174 | 0 |
| M13mp18/19 | 0 |
| pBR322 | 1 |
| pUC18/19 | 1 |
| pUC57 | 1 |
| pTZ19R/U | 1 |
| pBluescriptIIKS(-/+) | 1 |
| pBluescriptIISK(-/+) | 1 |
| pACYC177 | 1 |
| pACYC184 | 0 |
|
| Unit Definition | |
| One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA dcm--HindIII fragments in 1 hour at 37°C in 50ul of assay buffer. Assayed using lambda DNA dcm-, as one of two Eco31I recognition sites is difficult to cleave. |
|
| 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. |
