Dear all,
We use serum and or plasma as a matrix for molecular biology techniques. We do prepare serum or plasma with specific centrifugation related to each sample, then aliquoted samples were stored @-80°C. My question is, once thawed is it mandatory to proceed to an additional centrifugation (13000g, 10 minutes) of the serum or plasma samples? I know that I may loose material but I think this additional centrifugation may avoid any cellular contamination or cell debris contamination in the following step of my analyses.
Am I right to proceed to this additional "cleaning step" or not?
looking forward to having your return.
Best.
Arnaud
It is not mandatory. Whether or not it is advisable depends on your downstream analyses and the analytes you are looking at. In most cases, all you sediment are protein aggregates and cellular debris that appear after freeze-thaw. Therefore, you may not lose any of your relevant analytes and instead, end up having a cleaner starting material. If you are worried about losing material, just give your samples a quick spin (30 sec-2 min) in a small table top centrifuge instead of the long spin (13000g, 10 min). This should remove all aggregates and cell debris without affecting the downstream analyses.
- Freeze thawing can sometimes cause precipitation/denaturation and coagulation so if you have visible insoluble material in your serum my advice would be to always give it a quick 5 min spin and them remove the overlying liquid serum
Julie Joseph,
Thank you for your answer. I think I will proceed to a quick spin to be sure (and sure...) that I had removed all debris and protein contaminants..
Best
Laurence Stuart Dawkins-Hall,
Thank you for your advice, it goes with my thought to get the more pure & clean matrix to begin my experiments.
best.
Arnaud
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