I have certain cells that are contaminant cells in a suspension of cells. I found a buffer that selectively removes them, but a lot of protocols don't specify what volume one should use for that buffer.
For example - I have somatic cells contaminating sperm suspension, and buffer that removes them is composed of TritonX and SDS. How much buffer should I add - is it dependent on the concentration of cells, of volume of suspension pre-centrifugation, or is it irrelevant?
Generally it would be based on the number of cells you have. If your sample has contamination though is it better possibly to get a new sample than to remove the contamination.
About 5 to 10 volumes of your centrifuged pellet should be fine.
If you are unsure, you can do a pilot experiment by lysing the sample in a small volume. Based on the results you can dilute the sample or repeat with a larger volume.
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