Sdai Restriction Enzyme

E. Coli that carries the cloned sdaIR gene from Streptomyces diastaticus Ng 7-324

Supplied as a liquid in 10mM Tris-HCl, pH 7.5, 100mM KCl, 0.1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.

Molecular Biology Grade

10u/ul (lot specific)

Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

37°C

One unit is defined as the amount of Sdal required to digest 1 ug of lambda DNA in 1 hour at 37°C in 50ul of recommended reaction buffer.

10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM DTT, 0.2mg/ml BSA and 50% glycerol

MBS638383: Restriction Enzyme Buffer A, 10X: Dilute to 1X for use. 1X buffer compostion is 33mM Tris-acetate, pH 7.9 at 37°C, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.

MBS638379: Restriction Enzyme Buffer for ScaI, SdaI, 10X: Dilute to 1X for use. 1X buffer compostion is 37mM Tris-acetate, pH 7.0, 15mM Magensium acetate, 150mM potassium acetate, 0.1mg/ml BSA

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.

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, BseSI, Mph1103I, PstI, SduI

Number of Recognition Sites in DNA:
Lambda: 5
PhiX174: 0
pBR322: 0
pUC57: 0
pUC18/19: 1
pTZ19R/U: 1
M13mp18/19: 1

Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 5-fold overdigestion with Sdal (5 units/ug lambda DNA x 1 hour) (see Star Activity).

Ligation/Recutting Assay: The ligation and recleavage assay was replaced with L0 test after validating experiments showed L0 test ability to trace nuclease and phosphatase activities with sensitivity that is higher than L/R by a factor of 100.

Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded or double-stranded labeled oligonucleotide occurredduring incubation with 10 units of Sdal for 4 hours.

Blue/White (B/W) Cloning Assay: The B/W assay was replaced by the L0 test after validating experiments showed L0 test ability to trace nuclease and phosphatase activities with sensitivity that equals to that of B/W test.

Molecular Biology; MB-Enzymes; Restriction