BamH I is a restriction enzyme purified from Bacillus amyloliquefaciens H.
Reagents supplied: 10x BamHI and 10x EQ buffer
Unit definition: One unit is the amount of enzyme required to digest 1 μg of Lambda DNA in 60 min at 37oC in a total reaction volume of 50 μl.
Standard reaction conditions: In order to overcome DNA source variability, a ~10 fold overdigest is suggested:
10U BamH I
10x EQ buffer
Sterile ultrapure water
Up to 50 μl
Speed Digest reaction: Incubate for 15 minutes as in standard reaction in 1x BamHI or 1x EQ
Activity(%) in EnzyQuest buffers-Double digest tips
Reactions were performed as for unit definition reaction. The above data could be used as guideline for double digest reactions. Our enzymes are also active and compatible in buffer systems from other vendors.
Heat inactivation: 80oC for 20 minutes
dcm methylation: Not sensitiveCpG methylation: Not sensitive
Unit calculation assay conditions:
1x BamHI buffer
Lambda DNAIncubation at 37oC
50 mM KCl
10 mM Tris-HCl (pH 7.4)
0.1 mM EDTA
1 mM dithiothreitol
200 μg/ml BSA
Store at -20oC
Non-specific nuclease activity: 100 units of BamH I do not produce any unspecific cleavage products after 16 hrs incubation with 1 μg of Lambda DNA at 37oC.
Re-ligation and recut: After 20-fold overdigestion with BamH I, greater than 95% of the DNA fragments can be ligated and recut with this enzyme.
Star activity: Conditions of low ionic strength, high enzyme concentration, glycerol concentration >5% or pH >8.0 may result in star activity.
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