Answer :
Final answer:
We should not vortex the lysate because it can damage sensitive proteins, cause DNA shearing, and affect processes like lysis inhibition, therefore it is recommended to use gentler mixing methods and protease inhibitors for lysate processing.
Explanation:
We should not vortex the lysate because vortexing can be too harsh for the cell contents, leading to the potential damage or denaturation of the sensitive proteins such as the protein of interest. Proteases are released during cell lysis, which may start digesting proteins if not inhibited.
Vortexing can also introduce high shear forces that may cause DNA shearing, which could increase the viscosity of the lysate, making subsequent processing steps more difficult. Lysis inhibition is another consideration with certain organisms like phages where high extracellular phage concentrations inhibit lysis.
Therefore, gentler methods of mixing are often encouraged to preserve the integrity of the lysate fractions and ensure successful downstream applications such as fractionation. In summary, to maintain the quality and integrity of the lysate, it is crucial to proceed with less vigorous mixing and include protease inhibitors if necessary.
