Benzonase for Removal of Nucleic Acids in Protein Extraction

University of Birmingham
School of Biosciences
Postdoctoral Research Fellow

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Company:

EMD Millipore

Product Name:

Benzonase nuclease

Catalog Number:

70746-3

Separation of protein on chromatography columns from proteins extracted from gram negative bacteria treated with 1X BugBuster lysis reagent with Benzonase nuclease at 25 units/ml lysis reagent. Flow through the columns were efficient and got good yields of proteins.

Experimental Design and Results Summary

Application

Extraction of proteins from gram negative bacterial cells

Starting Material

Gram negative bacterial cell pellets (from 5 ml culture) treated with 1X Novagen BugBuster lysis reagent

Protocol Overview

I have used Novagen BugBuster protein extraction kit for extracting proteins. Followed protocol given in the kit's handbook. Used Benzonase nuclease with 1X Bugbuster lysis reagent at 25 units/ml. Then followed remaining steps for final separation of proteins using size exclusion column chromatography.

Tips

Dilute the benzonase nuclease enzyme in 50% glycerol if required to reduce the concentration.

Results Summary

After treating protein extracts with Benzonase nuclease, the viscosity was reduced to greater extent and flow through the chromatography columns were efficient.

DOI or PMID #

N/A

Additional Notes

None

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Summary

The Good

Easy to use and simple and efficient for removal contaminating nucleic acids.

The Bad

Bit expensive.

The Bottom Line

Ideal enzyme for protein preps and especially protein extracts to be separated on chromatographic columns.

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