|Molecular Biology||Storage: -20°CShipping: Blue Ice|
5'-C C T G C A^G G-3'
3'-G G^A C G T C C-5'
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.
E. coli that carries the cloned sdaIR gene from Streptomyces diastaticus Ng 7-324
R1625: Restriction Enzyme Buffer A, 10X
R1625-70: Restriction Enzyme Buffer for ScaI, SdaI, 10X
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.
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.
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.
Star Activity: An excess of enzyme
(7.5u/ug DNA x 1 hour) may result in star activity.
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:
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.