| 5'-A T G C A^T-3' |
|
| 3'-T^A C G T A-5' |
|
| Catalog # | M4680 |
| Source | Moraxella phenylpyruvica RFL1103 |
| Concentration | 10u/ul |
| Unit Definition | One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer. |
| R1625-04 (Included) | Restriction Enzyme Buffer E, 10X Incubate at 37°C |
| R1625 (Included) | Restriction Enzyme Buffer A, 10X |
| Provided to for double digests. |
| Diluent Buffer | 10mM Tris-HCl, pH 7.4 at 25°C, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. |
| Storage Buffer | 10mM Tris-HCl (pH 7.4 at 25°C), 200mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA,50% glycerol. |
| Enzyme Properties | Stability during Prolonged Incubation: A minimum of 0.3units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. |
| Methylation Effects | Dam/Dcm/CpG/EcoKl: never overlaps- no effect |
| EcoBl: may overlap-effect not determined |
| Thermal Inactivation | Enzyme is inactivated by incubation at 65°C for 20min. |
| Compatible Ends | Alw21I, PstI, SdaI, SduI |
| Number of Recognition Sites in DNA | Lambda: 14 |
| PhiX174: 0 |
| M13mp18/19: 0 |
| pBR322: 0 |
| pUC18/19: 0 |
| pUC57: 1 |
| pTZ19R/U: 0 |
| Storage and Stability | May be stored at 4°C for short-term only. For long-term storage, 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 buffe |
| Quality Control | |
| Overdigestion Assay | No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Mph1103I. |
|
| Ligation/Recutting Assay | After 50-fold overdigestion (3u/ug DNA x 17 hours) with Mph1103I, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.1uM. 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 with10units of restriction endonuclease for 4 hours. |
|
| Blue/White Cloning Assay | PUC57 was digested at a unique site with 10units of enzyme for 16 hours. After religation and transformation 0.3% of white colonies were detected. |
|
| 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. |
