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4 years ago
4 years ago by Hollie Fowler
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?
Found 1 matching solution for this experiment
|- Add lysozyme and Benzonase nuclease to the Qproteome Bacterial Protein Prep.
|- The cell pellets are washed twice with 20 mL of washing buffer (50 m Tris‐HCI, pH 7.2), centrifuged (14 000g, 30 min at 4°C), and then disrupted by an Qproteome Bacterial Protein Prep Kit (Qiagen, Hilden, Germany) according to the Qproteome Bacterial Protein Preparation Handbook.
|- The cell lysate supernatant is retained and then 100 mL of acetone are added and the samples are precipitated (12 h at −20°C). The proteins are pelleted (14 000g, 20 min) and resuspended in rehydration buffer (RB).
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