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You are here:Home » Molecular Biology » MB-Enzymes, Restriction » Bam HI

Bam HI

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Specifications

5'-G^G A T C C-3'
3'-C C T A G^G-5'
Catalog #B0070
Source Bacillus amyloliquefaciens H
Supplied withB0070A: Reaction Buffer (10X) for BamHI+, 10X
Supplied as a liquid in10mM Tris-HCl, pH 8.0, 5mM MgCl2, 100mM KCl, 0.02% Triton X-100, 0.1mg/ml BSA. Supplied with 2x1ml of Reaction Buffer (10X).
Concentration 10u/ul
Unit Definition One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA-Bsp120I fragments in 1 hour at 37°C in 50ul of assay buffer.
Diluent Buffer 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol.
Storage Buffer 10mM Tris-HCl, pH 7.4 at 25ºC, 200mM NaCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol.
Incubation Temp 37°C
Methylation EffectsBamHI does not cut GGATm4CC, GGATm5CC, GGAThm5Chm5C, GGAhm5UCC, but cuts GGATCm5C, GGm6ATCC, GGm6ATCm5C and GGATCm4C.
Number of Recognition Sites in DNA MoleculesLambda: 5
PhiX174: 1
M13mp18/19: 1
pBR322: 1
pUC18/19: 1
pUC57: 1
pTZ19R/U: 1
Stability during Prolonged Incubation A minimum of 0.5 units of enzyme are required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C.
Thermal Inactivation Only small amounts of enzyme (up to 10 units) can be inactivated at 80°C for 20min.
Compatible Ends BclI, BglII, Bsp143I, MboI, PsuI
Overdigestion Assay
No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with BamHI (see Star Activity).
Ligation/Recutting Assay
After 50-fold overdigestion (3u/ug DNA x 17 hours) with BamHI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.02uM. More than 95% of these can be recut.
Labeled Oligonucleotide (LO) Assay
No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of restriction endonuclease for 4 hours.
Blue/White Cloning AssayPUC57 was digested at a unique site with 10 units of enzyme for 16 hours. After religation and transformation 0.2% of white colonies were detected.
Digestion of Agarose-embedded DNA
A minimum of 5 units of enzume is required for complete digestion of 1ug of agarose- embedded lambda DNA in 16 hours.
Star Activity
Appears at > 5-fold overdigestion at (5u x 1h).


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