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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » Bpu10I

Bpu10I

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Specifications

5'-C C^T N A G C-3'
3'-G G A N T^C G-5'
Catalog #B2648
Concentration 5u/ul
Source E.coli that carries the cloned bpu10IR gene from Bacillus pumilus RFL10
Buffer 10mM Bis-Tris Propane-HCl (pH 6.5), 10mM MgCl2, 100mM KCl and 0.1mg/ml BSA. Incubate at 37°C.
Diluent Buffer 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. For longer periods–the Storage Buffer should be used.
Storage Buffer 10mM Tris-HCl (pH 7.5 at 25°C), 200mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Overdigestion AssayNo detectable change in the specific fragmentation pattern is observed after 48-fold overdigestion (3u/ug lambda DNA x 16 hours) with Bpu10I (see Star Activity).
Ligation/Recutting AssayAfter 50-fold overdigestion (3u/ug DNA x 17 hours) with Bpu10I, more than 90% of the pBR322 DNA fragments can be ligated at a 5'-termini concentration of 0.1uM. More than 90% of these can be recut. Where there is more than one Bpu10I recognition site in the substrate DNA, DNA fragments, obtained after cleavage with Bpu10I (due to its asymmetric recognition sequence), may join each other in 3 different ways and in only one case is the resulting sequence recognized by Bpu10I. Therefore, restriction after ligation yields no more than 30% efficiency. Other two sequences resulting from ligation of Bpu10I DNA fragments may be cleaved with Eco81I (SauI) and Bpu1102I (EspI)
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.
Star ActivityA large excess of enzyme (5u/ug DNA x 16 hours) may result in star activity.
Stability during Prolonged IncubationA minimum of 0.2units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Thermal InactivationEnzyme is inactivated by incubation at 80°C for 20min.
Compatible EndsBpu1102I, DdeI, Eco81I.
Number of Recognition Sites in DNA
Lambda19
PhiX1747
M13mp18/194
pBR3221
pUC18/190
pUC570
pTZ19R/U0
pBluescriptIIKS(-/+)0
pBluescriptIISK(-/+)0
pACYC1775
pACYC1843
NoteA large excess of enzyme (>4u/1ug DNA) may result in incomplete DNA cleavage. Therefore, we recommend increasing the incubation time instead of using an excess of Bpu10I.
Unit DefinitionOne unit is defined as the amount of enzyme required to digest 1ug of lambda DNA fragments in 1 hour at 37°C in 50ul of assay buffer.


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