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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » BstXI

BstXI

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Specifications

5'-C C A N N N N N^N T G G-3'
3'-G G T N^N N N N N A C C-5'
Catalog #B3001
SourceBacillus stearothermophilus X
Concentration10u/ul
Unit DefinitionOne unit is defined as the amount of BstXI required to digest 1ug of lambda DNA in 1 hour at 55°C in 50ul of assay buffer
Supplied WithR1625: Restriction BstXI Buffer A, 10X: Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA (pH 7.9 at 37°C).
R1625-03 Restriction BstXI Buffer D, 10X: Supplied as a liquid in 50mM Tris-HCl (pH 7.5), 10mM MgCl2, 100mM NaCl and 0.1mg/ml BSA. Incubate at 55°C. Incubation at 37°C results in 50% activity.
Diluent and Storage BufferSupplied as a liquid in 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.
Overdigestion Assay
No detectable change in the specific fragmentation pattern is observed after 15-fold overdigestion (15u/ug lambda DNA x 1 hours) (see Star Activity)
Ligation/Recutting Assay
After 50-fold overdigestion (3u/ug DNA x 17 hours) with BstXI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.13uM. 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 BstXI for 4 hours.
Blue/White Cloning Assay
The mix of pUC57/HindIII, pUC57/Eco32I and pUC57/PstI digests was incubated with 10 units of BstXI for 16 hours. After religation and transformation 0.1% of white colonies were detected.
Methylation Effects
DamNever overlaps- no effect
DcmMay overlap- blocked
CpGNever overlaps- no effect
EcpKl/EcoBlNever overlaps- no effect
Star Activity
Excess of BstXI (20u/ug DNA x 1 hour) may result in star activity.
Stability during Prolonged Incubation
A minimum of 0.1 units of BstXI is required for complete digestion of 1ug of lambda DNA in 16 hours at 55°C.
Thermal Inactivation
BstXI is inactivated by incubation at 65°C for 20min.
Digestion of Agarose-embedded DNA
A minimum of 5 units of BstXI is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Number of Recognition Sites in DNA
Lambda13
PhiX1743
pBR3220
pUC570
pUC18/190
pTZ19R/U0
M13mp18/190
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 12 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 diluent buffer.


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