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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » BspTI (AflII)

BspTI (AflII)

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Specifications

5'-C^T T A A G-3'
3'-G A A T T^C-5'
Catalog #B2970
Concentration 10u/ul
Source Bacillus species RFL1265I
Buffer (1X) (For 100% digestion)R1625-03: 50mM Tris-HCl (pH 7.5), 10mM MgCl2, 100mM NaCl and 0.1mg/ml BSA. Incubate at 37°C
Diluent BufferR1625: 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol, or a Storage Buffer.
FormSupplied as a liquid in 10mM Tris-HCl (pH 7.5 at 25°C), 200mM KCl, 1mM DTT, 0.1mM 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 BspTI.
Ligation/Recutting AssayAfter 50-fold overdigestion (3u/ug DNA x 17 hours) with BspLI, approximately 80% of the DNA fragments can be ligated at a 5'-termini concentration of 0.2uM. More than 90% of these can be recut.
Labeled Oligonucleotide (LO) AssayNo detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of restriction endonuclease for 4 hours.
Blue/White Cloning AssayThe mix of pUC57/HindIII, pUC57/PstI and pUC57/Eco32I digests was incubated with 10units of enzyme for 16 hours. After religation and transformation 0.3% of white colonies were detected.
Stability during Prolonged IncubationA minimum of 0.1units of enzyme is required for complete digestion of 1ug of DNA in 16 hours at 37°C.
Thermal InactivationEnzyme is inactivated by incubation at 65°C for 20min.
Digestion of Agarose-embedded DNAA minimum of 5units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Compatible EndsSmlI
Methylation Effects on Digestion
DamNever overlaps, no effect
DcmNever overlaps, no effect
CpGNever overlaps, no effect
EcoKINever overlaps, no effect
EcoBINever overlaps, no effect
Number of Recognition Sites in DNA
Lambda3
PhiX1742
M13mp18/190
pBR3220
pUC18/190
pUC570
pTZ19R/U0
pBluescriptIIKS(-/+)0
pBluescriptIISK(-/+)0
pACYC1770
pACYC1840
Unit DefinitionOne unit is defined as the amount of enzyme required to digest 1ug of lambda DNA dam-in 1 hour at 37°C in 50ul of assay buffer.


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