|Molecular Biology||Storage: -20°CShipping: Blue Ice|
5'-G^G A T C C-3'
3'-C C T A G^G-5'
Source: Bacillus amyloliquefaciens H
One unit is defined as the amount of Bam HI required to digest 1ug of lambda DNA-Bsp120I fragments in 1 hour at 37°C in 50ul of assay buffer.
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 37°C).
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.
10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.
Supplied as a liquid in 10mM Tris-HCl, pH 7.4 at 25ºC, 200mM NaCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 50% glycerol.
Incubation Temp: 37°C
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 37°C.
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.
BclI, BglII, Bsp143I, MboI, PsuI
Number of Recognition Sites in DNA:
pBR322, pUC57, pUC18/19, pTZ19R/U,
Bam HI (up to 10 units) is inactivated by incubation at 80°C for 20min.
Quality Control Assay Data:
No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours).
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 -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.