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Pierce™ Universal Nuclease for Cell Lysis, 5 kU - FAQs

Pierce™ Universal Nuclease for Cell Lysis, 5 kU - FAQs

View additional product information for Pierce™ Universal Nuclease for Cell Lysis - FAQs (88702, 88701, 88700)

3 product FAQs found

At which concentration/dilution should the Pierce Universal Nuclease for Cell Lysis (Cat. No. 88700) be used, and what is the suggested incubation time and incubation temperature?

We would suggest the following procedure (note that you should determine yourself empirically the optimal incubation time, incubation temperature and also dilution): Please add 25 U or 0.1 µL Pierce Universal Nuclease for Cell Lysis (Cat. No. 88700) to 1 mL of cell lysate (dilution 1:10,000) and incubate at room temperature for 10-15 minutes. If you want to dilute the Nuclease for facilitating pipetting, please do this with Tris- or PBS-buffer at pH 7-8.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

After cell lysis, my mass spectrometry sample is very viscous and difficult to pipette. How do I reduce the sample viscosity?

High sample viscosity after lysis is due to release of DNA from the nucleus. Sonication or addition of a nuclease such as the Pierce Universal Nuclease (Cat. No. 88700, 88701, or 88702) can be used to degrade DNA and reduce sample viscosity.

Find additional tips, troubleshooting help, and resources within our Mass Spectrometry Support Center.

Can you explain how the B-PER Bacterial Protein Extraction Reagent lyses cells?

The B-PER Reagent solution contains a proprietary, mild, non-ionic detergent in 20 mM Tris-HCl, pH 7.5. It effectively disrupts cells and solubilizes native or recombinant proteins without denaturation. The reagent creates holes in the cell membrane that will leak out cytosolic proteins. The sample may become very viscous when the bacterial chromosome is released. We recommend adding DNAse I (Cat. No. 90083) to the reagent to reduce viscosity. For better lysis efficiency and if there are inclusion bodies, we recommend adding Lysozyme (Cat. No. 90082) to the reagent. Alternatively, you may purchase the B-PER Bacterial Protein Extraction Reagent with Enzymes Kit (Cat. No. 90078 or 90079) that includes the B-PER Bacterial Protein Extraction Reagent, DNase I, and Lysozyme.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.