Enzymes that use DNA as a substrate
Bam HI is a restriction endonuclease and that means it cleaves the DNA in a specific sequence of nucleotides. Was isolated from a microorganism, more precisely from Bacillus amyloliquefaciens. This enzyme goes through all DNA molecule and then, when recognizes the sequence GGATCC and cleaves the molecule between the two guanine nucleotides.
Bam HI is more effective at 37ºC and isn’t sensitive to heat inactivation. It is storage at -20ºC in this conditions: (10 mM Tris-HCl; 50 mM KCl; 1 mM DTT; 0.1 mM EDTA; 200 μg/ml BSA; 50% Glycerol; pH 7.4 @ 25°C). Also, Bam HI can cut substrate DNA in 5-15 minutes and can be digested overnight without destroying the DNA.
The principal applications of Bam HI are molecular cloning, restriction site mapping, Genotyping, Restriction fragment length polymorphism (RFLP) and single nucleotide polymorphism (SNP).
The buffer used to incubate the Bam HI is different from company to company, but one of them is: (100 mM NaCl;50 mM Tris-HCl;10 mM MgCl2;100 μg/ml BSA ;(pH 7.9 @ 25°C))
References:
New England Biolabs.inc:
ThermoFisher scientific:
Protein sequence analysis and classification:
Grupo2.3: Sofia Salgado, Simão Ferreira, Tânia Castro, Tiago Ribeiro, Eduardo Faia
Applied Biology 2nd year, University of Minho
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