I would like to use DNase I (from bovine pancreas, Roche) to get rid of DNA. Roche/Sigma does some recommendations (1 mg/ml enzyme, pH 7.0, Ca 0.12 mM, some Mg), but I did not find a comprehensive buffer recipee. NEB suggests 10 mM Tris-HCl pH 7.6, 2.5 mM MgCl2, 0.5 mM CaCl2. Do you have any recommendation?
The DNA to digest is a possible contamination on the metal beads we use for nucleic acid isolation. The plan is to incubate the beads with DNase I, wash, and then autoclave before use.
The DNase concentration is so high that optimization is hardly necessary. Mg2+ is essential.
The DNase concentration is so high that optimization is hardly necessary. Mg2+ is essential.
Hi,
The RNAse free DNAse I from Roche for applications in RT-PCR tempates is supplied along with a buffer that also contains Mg2+...See if you can get complete buffer composition in the available literature (online)
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