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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » Bsh1236I (FnuDII)

Bsh1236I (FnuDII)

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Specifications

5'-C G C G-3'
3'-G C G C-5'
Catalog #B2922
SourceBacillus sphaericus RFL1236
Concentration 10u/ul
Incubation Temperature 37°C
Storage BufferSupplied as a liquid in 10mM Tris-HCl, pH 7.5, 100mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 50% glycerol.
Diluent Buffer10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
Supplied withR1625: Restriction Enzyme Buffer A, 10X: 33mM Tris-acetate, pH 7, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA
R1625-04 Restriction Enzyme Buffer E, 10X, 10mM Tris-HCl, pH 8.5 at 37°C, 10mM MgCl2, 100mM KCl, 0.1mg/ml BSA
Storage and StabilityAliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Enzyme Properties
Methylation Effect
Blocked by CpG methylation.
Dam, Dcm, EcoKl, EcoBlNeber overlaps, no effect
Stability during Prolonged Incubation
A minimum of 0.1units of B2922 is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Thermal Inactivation
B2922 is inactivated by incubation at 65°C for 20min.
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 Bsh1236I.
Ligation/Recutting Assay
After 50-fold overdigestion (3u/ug DNA x 17 hours) with Bsh1236I, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 1.6uM. 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 B2922 for 4 hours.
Number of Recognition Sites in DNA
Lambda157
PhiX17414
M13mp18/1918
pBR32223
pUC18/1910
pUC5711
pTZ19R/U11
Protocol for Digestion
Add
Nuclease free water16ul
R1625-042ul
DNA (0.5-1ug/ml)1ul
B29220.5-2ul
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
R1625-042ul
B29221-2ul
Mix gently and spin down for a few seconds.
Incubate at 37ºC for 1-16 hours.


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