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

BglII

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Specifications

5'-A^G A T C T-3'
3'-T C T A G^A-5'
Catalog #B1087-05
Enzyme PropertiesMethylation Effects:
Dam: completely overlaps-no effect.
Dcm/CpG/EcoKl: never overlaps-no effect.
EcoBl: may overlap-cleavage impaired.
Stability during Prolonged IncubationA minimum of 0.1 units of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Thermal InactivationEnzyme is not inactivated by incubation at 80°C for 20 minutes.
Digestion of Agarose-embedded DNAA minimum of 5 units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours.
Compatible EndsBamHI, BclI, Bsp143I, MboI, PsuI
Number of Recognition Sites in DNALambda: 6
PhiX174: 0
M13mp18/19: 1
pBR322: 0
pUC18/19: 0
pUC57: 0
pTZ19R/U: 0
Concentration10u/ul
SourceBacillus globigii
Form;
Supplied as aliquid in 50mM Tris-HCl pH 7.5, 10mM MgCl
Diluent Buffer
For short term storage (3-4 weeks, dilute with Diluent Buffer (10mM Tris-HCl, pH 7.4,,100mM potassium chloride, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol).
Storage Buffer
For long term storage, use Storage Buffer (10mM Tris-HCl, pH 8.2, 200mM potassium chloride, 1mM DTT, 0.1mM EDTA, 0.5mg/ml BSA, 50% glycerol).
Quality Control
Overdigestion Assay
No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with BglII.
Ligation/Recutting Assay
After 50-fold overdigestion (3u/ug DNA x 17 hours) with BglII, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.06uM. 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 10units of BglII for 4 hours.
Blue/White Cloning Assay
The mix of pUC57/HindIII, pUC57/PstI and pUC57/Eco32I digests was incubated with 10units of BglII for 16 hours. After religation and transformation <1% of white colonies were detected.
Protocol for Digestion
Add
Nuclease free water16ul
R1625-032ul
DNA (0.5-1ug/ml)1ul
B1087-050.5-2ul
Mix gently and spin down for a few seconds. Incubate at 37ºC for 1-16 hours.
Protocol for Digestion Directly after Amplification
Add
PCR Reaction Mixture10ul (~0.1-0.5ug of DNA)
Nuclease free water18ul
R1625-032ul
B1087-051-2ul
Mix gently and spin down for a few seconds.
Incubate at 37ºC for 1-16 hours.


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