Technical Data
X0960
XagI (EcoNI)
1000U
Molecular Biology Storage: -20CShipping: Blue Ice
5'-C C T N N^N N N A G G-3'
3'-G G A N N N^N N T C C-5'

Source:
Xanthobacter agilis Vs18-132

Supplied With:
R1625: Restriction Enzyme Buffer A, 10X; (1X compostition after dilution)-33mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.

R1625-04: Restriction Enzyme Buffer E, 10X: 10mM Tris-HCl, pH 8.5,10mM magnesium chloride, 100mM potassium chloride, 0.1mg/ml BSA

Concentration:
10u/ul

Unit Definition:
One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer.

Diluent Buffer:
10mM Tris-HCl (pH 7.4 at 25C), 100mM potassium chloride, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.

Storage Buffer:
10mM Tris-HCl (pH 7.5 at 25C), 100mM potassium chloride, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 320-fold overdigestion (20u/ug lambda DNA x 16 hours) with XagI.

Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with XagI, approximately 80% of the DNA fragments can be ligated in the reaction mixture containing 2040u of T4 DNA Ligase/1ug of fragments and 10% of PEG at a 5'-termini concentration of 0.1M. 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: The mix of pUC57/HindIII, pUC57/Eco32I and pUC57/PstI digests was incubated with 10 units of enzyme for 16 hours. After religation and transformation 0.2% of white colonies were detected.

Stability during Prolonged Incubation: A minimum of 0.1 units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C.

Thermal Inactivation: Enzyme is inactivated by incubation at 65C for 20min.

Digestion of Agarose-embedded DNA: A minimum of 5 units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.

Number of Recognition Sites in DNA:
Lambda: 9
PhiX174: 0
M13mp18/19: 0
pBR322: 1
pUC18/19: 0
pUC57: 0
pTZ19R/U: 0
pBluescriptIIKS(-/+): 0
pBluescriptIISK(-/+): 0
pACYC177: 1
pACYC184: 1

Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.