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Work Faster be More Accurate, biology Laboratory Organization Tip Different Cell Disruption Methods Pipette Eppi Pen
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Work Faster 🐝 More Accurate
Lab Organization Tip
Different Cell Disruption Methods
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Work Faster 🐝 More Accurate
If you use certain chemicals regularly, prepare concentrate stock solutions that you can later add to your samples. It reduces deviations between different experiments.
How? You can not prepare a buffer 100% the same each time.
⚠ Remember: If you add your stock solution to the buffered sample it will diluted if you don’t prepare it in the same buffer.
Lab Organization Tip
Assign different colors to different areas in your lab. Mark with a colored dot the samples, chemicals, etc.
Different Cell Disruption Methods
Physical Methods
🟡 Sonication must be used w/ ear protection on ice/in the cold room. Try to pulse it to avoid foaming.
🟡 Beads mill gets hot as well.
🟡 Passing through a syringe for sensitive mammalian cells.
Chemical Methods
🟡 RIPA Lysis Buffer is common
One of the ingratiates of RIPA buffer is the detergent SDS. You can use other detergents. However, detergents might desaturate your proteins.
🟡 ddH2O: Some sensitive mammalian cells (Erythrocytes PMID: 20234842) can burst due to osmatic pressure. The cells have a higher slat conc and thus swell until they burst open. Make sure to wash the cells first clean of buffer residues. Other more resilient cells can withsdant ddH20. (PMID: 25870207)
Enzymatic Methods
🟡 Some enzymatic kits can provide a sensitive cell lysis. If you try to isolate nucleic acids (RNA/DNA), proteases degrade RNases and DNases.
Con; expensive for large scale!
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