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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » XapI (ApoI)

XapI (ApoI)


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5'-Pu^A A T T Py-3'
3'-Py T T A A^Pu-5'
Catalog #X0985
SourceXanthomonas ampelina Slo 51021
FormSupplied as a liquid (Storage Buffer) in 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Restriction Enzyme Buffer A, 10X (for 100% digestion) R162533mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate and 0.1mg/ml BSA. Incubate at 37°C.
Unit DefinitionOne unit is defined as the amount of XapI required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer..
Storage and StabilityMay 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 15-fold overdigestion with XapI(15u/ug lambda DNA x 1 hours) .
Ligation/Recutting Assay
After 10-fold overdigestion (5u/ug DNA x 2 hours) with HpaII, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.2uM. More than 95% of these can be recut.
Labeled Oligonucleotide (LO) AssayNo detectable degradation of single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of XapI for 4 hours.
Blue/White Cloning Assay
pUC57 was digested at a unique site with 10 units of XapI for 1 hours. After religation and transformation 0.2 % of white colonies were detected.
Star ActivityAn excess of XapI (20u/ug DNA x 1 hour) may result in star activity.
Stability during Prolonged IncubationA minimum of 0.1 units of XapI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Digestion of Agarose-embedded DNAA minimum of 5 units of XapI is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Compatible Ends
EcoRI, MunI, TasI
Number of Recognition Sites in DNA
Lambda58PhiX174: 5
Protocol for Digestion
Add44ul nudlease free dH2O
5ul Restriction Enzyme Buffer A, 10X
1ul DNA (0.5-1ug/ul)
0.5-2ul XapI
Mix gently and spin down for a few seconds.
Incubate at 3ºC for 1-2 hours.
Protocol for Digestion of PCR products Directly after Amplification
Add10ul (~1ug DNA)
16ul nuclease free dH2O
2ul Restriction Enzyme Buffer A, 10X
1-2ul XapI
Mix gently and spin down for a few seconds.
Incubate at 3ºC for 1-16 hours.
Thermal Inactivation
XapI is inactivated by incubation at 65°C for 20min.

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