| 5'-C C T G C A^G G-3' |
|
| 3'-G G^A C G T C C-5' |
|
| Catalog # | S0549 |
| Concentration | ~10u/ul |
| Unit Definition | One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA fragments in 1 hour at 37°C in 50ul of assay buffer. |
| Source | E. coli that carries the cloned sdaIR gene from Streptomyces diastaticus Ng 7-324 |
| Supplied With | R1625: Restriction Enzyme Buffer A, 10X |
| R1625-70: Restriction Enzyme Buffer for ScaI, SdaI, 10X |
| Diluent Buffer | For short term storage (~3-4 weeks)-10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. For longer periods–the Storage Buffer should be used. |
| Storage Buffer | 10mM Tris-HCl (pH 7.5 at 25°C), 100mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol. |
| 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 buffer. |
| Quality Control | |
| Overdigestion Assay | No detectable change in the specific fragmentation pattern is observed after 5-fold overdigestion (5 units/ug lambda DNA x 1 hour) with SdaI (see Star Activity). |
|
| Ligation/Recutting Assay | After 5-fold overdigestion (2.5 units/ug DNA x 2 hours) with SdaI, 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. |
|
| Blue/White Cloning Assay | PUC19 DNA was digested at a unique site with 10 units of enzyme for 16 hours. After religation and transformation 0.1% of white colonies were detected. |
|
| Enzyme Properties | |
| Star Activity | An excess of enzyme |
| (7.5u/ug DNA x 1 hour) may result in star activity. |
|
| Methylation Effects | |
| Dam/Dcm/CpG/EcoKl/EcoBl | Never overlaps - no effect. |
|
| 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 complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours. |
|
| Compatible Ends | Alw21I, Mph1103I, PstI, SduI |
|
| Number of Recognition Sites in DNA | |
| Lambda | 5 |
| PhiX174 | 0 |
| M13mp18/19 | 1 |
| pBR322 | 0 |
| pUC18/19 | 1 |
| pUC57 | 0 |
| pTZ19R/U | 1 |
|
| 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. |
