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BamHI is a restriction enzyme isolated from Bacillus amyloliquefaciens that cleaves the palindromic sequence G▼GATCC.

Concentration: 10u/µL
Source: Bacillus amyloliquefaciens H
Reagents supplied with: The Enzyme comes with the Unique BamHI Buffer (recommended buffer) and the universal Sipmle Buffer for double digests. All our restriction buffers already contain BSA for your convenience. The enzyme is fully active in the respective buffers from other suppliers.
Store at: -20 °C

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Unit definition: One unit is defined as amount of enzyme required to produce a complete digest of 1 µg Lambda DNA in 60 min in a total reaction volume of 50µL under the assay conditions.
Unit substrate: Lambda DNA.
Unit calculation assay conditions: 100 mM NaCl, 10 mM Tris-HCl (pH 7.9 at 25°C), 5 mM MgCl2, 1 mM dithiothreitol, 100 μg/ml BSA. Incubate at 37 °C.
Absence of contaminants: 100 units of BamHI incubated for 16 hours at 37 °C with 1 μg of λ DNA resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis. After 50-fold overdigestion with BamHI, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.
Storage buffer: BamHI is supplied in 50 mM KCl, 10 mM TrisHCl (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 200 μg/ml BSA and 50% glycerol.
Inactivation: Heat to 80 °C for 20 minutes. Inactivation is also possible by addition of EDTA to a final concentration of 10mM or SDS to a concentration of 0.1 %. Alternatively use our environmentally friendly 6x orange gel loading buffer. For further enzymatic reactions remove SDS or EDTA by spin column-based PCR purification kit.
Methylation Sensitivity: dam methylation: Not sensitive dcm methylation: Not sensitive CpG methylation: Not sensitive
Star activity: Conditions of low ionic strength, high enzyme concentration, glycerol concentration >5% or pH>8.0 may result in star activity.

Activity in Simplebiotech buffers:

Activity in Simplebiotech buffers:

L M H SH Simple 2x Simple
50-75 75-100 100   50-75  100 50-100


*May exhibit star activity under these conditions.

Nuclease free water             x µL
10x Unique BamHI buffer      2 µL
DNA (0,5-1µg)                     1 µL
10 U BamHI                         1 µL
                                 Final volume 20µL

Incubate at 37 °C for 15min. Restriction enzymes are stored in 50% glycerol. The glycerol concentration in the reaction mix should be kept below 10%. When digesting more than 1µg DNA, scale up the restriction mix or prolong the restriction digest.




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