|Molecular Biology||Storage: -20°CShipping: Blue Ice|
5'-A G T^A C T-3'
3'-T C A^T G A-5'
Source: Streptomyces caespitosus
R1625-70: Restriction Enzyme Buffer for ScaI, SdaI, 10X
37mM Tris-acetate (pH 7.0), 15mM magnesium acetate, 150mM potassium acetate and 0.1mg/ml BSA. Incubate at 37°C
Diluent Buffer: 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), 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.5mg/ml BSA and 50% glycerol.
Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 5-fold overdigestion (5u/ug lambda DNA x 1 hour) with ScaI (see Star Activity).
Ligation/Recutting Assay: After 5-fold overdigestion (5u/ug DNA x 1 hour) with ScaI, more than 80% of the DNA fragments can be ligated at a 5'-termini concentration of 0.05uM. More than 90% 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 (7.5u/ug DNA x 1 hour) may result in star activity.
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: Only small amounts of enzyme (up to 10 units) can be inactivated by incubation at 80°C for 20min.
Digestion of Agarose-embedded DNA: A minimum of 20 units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Number of Recognition Sites in DNA:
5 0 0 1 1 1 1 1 1 1 1
* Unit Definition. One unit is defined as the amount of enzyme required to digest 1µg of lambda DNA in 1 hour at 37°C in 50µl of assay buffer.
NR: buffer is not recommended, since star activity appears at 5-fold overdigestion (5 units x 1 hour).
Product Use Limitation
Home Search Contacts Order Catalog Support
Updated March 07, 2002 15:46