Technical Data
B0070
Bam HI
10,000u
Molecular Biology Storage: -20CShipping: Blue Ice
5'-G^G A T C C-3'
3'-C C T A G^G-5'

Source: Bacillus amyloliquefaciens H

Concentration: 10u/ul

Unit Definition:
One unit is defined as the amount of Bam HI required to digest 1ug of lambda DNA-Bsp120I fragments in 1 hour at 37C in 50ul of assay buffer.

Supplied With:
R1625: Restriction Enzyme Buffer A, 10X: Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA (pH 7.9 at 37C).
B0070A Reaction Buffer, 10X: Supplied as a liquid in 10mM Tris-HCl, pH 8.0, 5mM MgCl2, 100mM KCl, 0.02% Triton X-100, 0.1mg/ml BSA. Dilute and use at 1X.

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

Storage Buffer:
Supplied as a liquid in 10mM Tris-HCl, pH 7.4 at 25C, 200mM NaCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 50% glycerol.

Incubation Temp: 37C
Enzyme Properties:
Methylation Effects:
Dam: completely overlap - blocked
Dcm, CpG: May overlap- no effect
EcoKl, EcoBl: Never overlaps- no effect

Stability during Prolonged Incubation:
A minimum of 0.5 units of Bam HI is required for complete digestion of 1ug of lambda DNA in 16 hr at 37C.

Digestion of Agarose-embedded DNA:
A minimum of 5 units of Bam HI is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hr.

Compatible Ends:
BclI, BglII, Bsp143I, MboI, PsuI

Number of Recognition Sites in DNA:
Lambda: 5
PhiX174: 0
pBR322, pUC57, pUC18/19, pTZ19R/U,
M13mp18/19: 1

Thermal Inactivation:
Bam HI (up to 10 units) is inactivated by incubation at 80C for 20min.

Quality Control Assay Data:
Overdigestion Assay:
No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours).

Ligation/Recutting Assay:
The ligation and recleavage assay was replaced with LO test after validating experiments showed LO 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 10 units of Bam HI for 4 hr.

Blue/White Cloning Assay:
The B/W assay was replaced with LO test after validating experiments showed LO test ability to detect nuclease and phosphatase activities with sensitivity that equals to that of B/W test.

Storage and Stability:
For long-term storage, 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.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.