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Technical Data |
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S0549 |
SdaI (Sse8387I) |
300u 1500u |
| Molecular Biology | Storage: -20°CShipping: Blue Ice |
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5'-C C T G C A^G G-3' 3'-G G^A C G T C C-5' 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. |
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