In article <bpmurray*STUFFER*-0801991828040001 at mac-daddy.ucsf.edu>,
bpmurray*STUFFER*@socrates.ucsf.edu (Bernard P. Murray, PhD) wrote:
>> Is this the same thing as eg. micrococcal nuclease?
> How do you inactivate benzonase? Can you treat a
> protein sample and then inactivate the enzyme so
> that you can re-introduce DNA or RNA and not have
> them chewed up? (ie. would it be any use in cleaning
> samples for transcription/translation?)
> Thanks for any (scientific) info,
> Bernard
> --
> Bernard P. Murray, PhD
> Dept. Cell. Mol. Pharmacol., UCSF, San Francisco, USA
>
Dear Bernard,
Benzonase is not identical with a micrococcal nuclease. Benzonase is a trade
name for a recombinant Serratia marcescens endonuclease. The inactivation of
the enzyme affords rather harsh conditions i.e. 70°C, >0.02 NaOH, 15 min.
That means that this kind of irreversible inactivation can hardly be used
without damaging your target molecule too. No specific inhibitors are known
for the enzyme. The removal is usually done by classical downstream
processing (ultrafiltration, size exclusion chromatography, IEX or HIC)
Please note: Never use Benzonase in a lab where you like to isolate DNA/RNA.
The enzyme is designed for downstream processing of recombinant proteins,
especially in the biopharmaceutical industry, where DNA/RNA has to be
completely removed from the final product and where viscosity caused by
nucleic acids is a problem. Frank
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