Technical Data
B2645
Bpl I
100U
500U
Molecular Biology Storage: -20CShipping: Blue Ice
5'-^ 8(N) G A G (N)5 C T C (N)13^-3'
3'-^13(N) C T C (N)5 G A G (N) 8^-5'

BplI requires only Mg2+ for its activity, but is stimulated by S-adenosylmethionine. 0.05mM S-adenosylmethionine gives more than a 100-fold increase of BplI activity. The cleavage of DNA by BplI is never complete.

Source:
Bacillus pumilus

Concentration:
5 units/ul

Unit Definition:
One unit is defined as the amount of Bpl I required to digest 1ug of lambda DNA-Xhol in 1 hour at 37C in 50ul of assay buffer (a maximal cleavage level is achieved at which no change in the fragmentation pattern is observed with further increase of enzyme).

Storage Buffer:
Supplied as a liquid in 10mM Tris-HCl, pH 7.5, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol.

Buffers Supplied:
R1625 Restriction Enzyme Buffer A, 10X:
33mM Tris-acetate, pH 7, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/mlBSA
B2645-01 SAM
0.05mM S-adenosylmethionine

Storage and Stability:
Aliquot to avoid repeated freezing and thawing. Store at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Stability during Prolonged Incubation:
A minimum of 0.3 units of Bpl I is required for complete digestion of 1ug of DNA in 16 hours at 37C.

Thermal Inactivation:
Bpl I is inactivated by incubation at 65C for 20min.

Methylation Effect:
DAM, DCM, EcoKl, EcoBl: never overlaps - no effect
CpG: may overlap - no effect

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

Digestion of Agarose-embedded DNA:
A minimum of 10 units of the Bpl I is required for complete digestion of 1ug of agarose-embedded lambda DNA in 16 hours.

Overdigestion Assay:
No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with BplI.

Ligation/Recutting Assay:
After 10-fold overdigestion (0.6u/ug DNA x 17 hours) with BplI, more than 70% of the DNA fragments can be ligated at a 5'-termini concentration of 0.03M. None of these can be recut due to the methylation at the recognition sequence by Bpll.

Labeled Oligonucleotide (LO) Assay:
No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of Bpl I for 4 hours.

Blue/White Cloning Assay:
The mix of pUC57/HindIII, pUC57/PstI and pUC57/Eco32I digests was incubated with 10units of Bpl I for 16 hours. After religation and transformation the background level of white colonies was <1%.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.