BamHI

Beschreibung

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:

*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.