How can I deal with my pellet being too viscous?

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5 years ago

Hollie Fowler

2

How can I deal with my pellet being too viscous?

I am using 8M Urea to lyse my cells but after centrifugation my pellet is very viscous. This causes me trouble when I try to collect the supernatant. Is there any way to avoid it and do you think it will have an effect on the concentration of my protein?

Experiment: Protein isolation Bacteria - Clostridium difficile

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**Dr. Tushar Tomar** · Answer 1 · 2019-05-09T14:42:11+00:00

This usually occurs because of the DNA that is released after lysing the cells. You can try to shear your sample by running it up and down. Alternatively, the most effective thing you can do is to sonicate your sample a few times (5-10 times) in small intervals of 1-5 seconds. Make sure to keep the sample on ice-cold water at all times when sonicating it. Do not forget to clean the sonication probe tip with ethanol and MQ water while changing the samples.