Technical Data
BsuRI (HaeIII)
Molecular Biology Storage: -20CShipping: Blue Ice
5'-G G^C C-3'
3'-C C^G G-5'


Bacillus subtilis R

Diluent Buffer:
10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.

Thermal Inactivation:
BsuRI is inactivated by incubation at 80C for 20 minutes.

Storage Buffer:
10mM Tris-HCl, pH 7.5, 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA and 50% glycerol.

Supplied with:
R1625-Restriction Enzyme Buffer A, 10X: Dilute to 1X for use. 1X buffer composition is 33mM Tris-acetate (pH 7.9 at 37C), 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 37C

Incubation Temperature: 37C

Unit Definition:
One unit is defined as the amount of BsuRI required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer.

Methylation Effects:
Dam, EcoKl,EcoBl: never overlaps - no effect.
Dcm, CpG: may overlap - no effect.

Protocol for Digestion:
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 37C for 1-16 hours.

Protocol for Digestion Directly after Amplification:
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 37C for 1-16 hours.

Storage and Stability:
Aliquot and store at -20C. Aliquots are stable for 6 months after receipt at -20C. 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) Assay: No 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 37C.

Number of Recognition Sites in DNA:
Lambda: 149
PhiX174, pUC18/19: 11
M13mp18/19: 15
pBR322: 22
pUC57: 13
pTZ19R/U: 12
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.