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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » BsuRI (HaeIII)

BsuRI (HaeIII)

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Specifications

5'-G G^C C-3'
3'-C C^G G-5'
Catalog #B3100
Concentration
10u/ul
SourceBacillus subtilis R
Diluent Buffer10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
Thermal InactivationBsuRI is inactivated by incubation at 80°C for 20 minutes.
Storage Buffer10mM Tris-HCl, pH 7.5, 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Supplied withR1625-Restriction Enzyme Buffer A, 10X: Dilute to 1X for use. 1X buffer composition is 33mM Tris-acetate (pH 7.9 at 37°C), 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA.
R1625-04 Dilute to 1X for 100% BsuRI digestion. 10mM Tris-HCl, pH 8.5, 10mM MgCl2, 100mM KCl and 0.1mg/ml BSA. Incubate at 37°C
Incubation Temperature 37°C
Unit DefinitionOne unit is defined as the amount of BsuRI required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer.
Methylation EffectsDam, EcoKl,EcoBl: never overlaps - no effect.
Dcm, CpG: may overlap - no effect.
Protocol for DigestionAdd:
Nuclease free water: 16ul
R1625-04: 2ul
DNA (0.5-1ug/ml): 1ul
B3100: 0.5-2ul
Mix gently and spin down for a few seconds. Incubate at 37ºC for 1-16 hours.
Protocol for Digestion Directly after AmplificationAdd:
PCR Reaction Mixture: 10ul (~0.1-0.5ug of DNA)
Nuclease free water: 18ul
R1625-04: 2ul
B3100: 1-2ul
Mix gently and spin down for a few seconds.
Incubate at 37ºC for 1-16 hours.
Storage and StabilityAliquot and store at -20°C. Aliquots are stable for 6 months after receipt 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 assay buffer.
Overdigestion Assay
No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with BsuRI.
Ligation/Recleavage (L/R) Assay
The ligation and recleavage assay was replaced with L0 test after validating experiments showed L0 test ability to trace nuclease and phosphatase activities with sensitivity that is higher than L/R by a factor of 100.
Labeled Oligonucleotide (LO) AssayNo detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of BsuRI for 4 hours.
Stability during Prolonged Incubation
A minimum of 0.1units of BsuRI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Number of Recognition Sites in DNA
Lambda149
PhiX174, pUC18/1911
M13mp18/1915
pBR32222
pUC5713
pTZ19R/U12


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