Hello everyone,
I need exosome free serum for my cell culture media. I`d tried to use Beckman Coulter - Optima L-100 XP at 100k g but my tubes (Thinwall, Polyallomer, 94 mL, 38 x 102 mm) got crushed and leaked even in 20 minutes. I wonder if I should use another type of tube? I don't know where did I do wrong...
Recently, I used the Beckman ultracentrifuge, it crushed and leaked too, the engineer suggested me to use another type tube, and he could not give me a satisfied explanation.
Xiaoyun Li so, could you please share the reorder code of this tube? Thank you.
Hi,
Could you please provide some information about the rotor and tubes in detail?
I routinely harvest exosome depleted FBS using polyallomer tubes in SW-28 rotor and never faced such a problem before.
However, I faced similar problem earlier with polycarbonate tubes.
Such problems can arise due to imbalanced tubes or unclean rotor which can cause cracks in the tube.
I assume you balance the tubes prior to centrifugation.
Follow the standard exosome isolation protocols described in the literature for ultracentrifugation, but discard exosomes and preserve the supernatant. "Tougher" tubes are important. Backup: exosome depleted FBS (Gibco) which is "ready to go"
Vasu Saini thank you for your answer. I balanced the tubes carefully. My rotor is 45 Ti and we used polyallomer tubes too. Your rotor is different so maybe these tubes can be proper to it. But the firm says polyallomer tubes are suitable for my rotor, I do not understand the problem.
Alexander Sasha Vlassov thank you for answer. Unfortunately, I am at the starting point for these protocols. Because my problem is about ultracentrifugation process. I know about exo-free FBS which ready to use but my budget is not enough to afford this product...
One possible problem could be that your tubes are not compatible with your rotor.
Assuming that you have been using compatible tubes as mentioned in the catalog of rotor. You can try to lower down the volume of FBS which you use per tube.
Other alternative is to use thickwall polyallomer tubes which are more tough than thinwall tubes. I wouldn't recommend using polycarbonate tubes as they are more prone to get cracks due to dust or deformities inside the rotor.
Vasu Saini thank you for your answers. Next time I will consider these informations.
Hi Gülşah Torkay ,
Were you able to resolve the issue in this rotor? Additional considerations are making sure the tube is properly supported (i.e., using the required caps and filling the tube as full as possible).
Please let me know if you have additional questions.
Best,
Ross
Hi Ross VerHeul
I think you are right about that the problem may be caused by tubes and caps. That was my first time in use and I was not sure about the tubes are suitable for liquid or not. Because this device is not belonging to my university, I have not tried to use it again. I will use after be sure. Thank you!
Did you put the aluminium red cap on the tube when spinning them or you let them open?
Jorgaq Pata the caps were blue as I remember. There was a tool for closing them.
Normally the cap for these tubes is red, the blue is for the thickwall tubes, I don't know whether this could have played any role in your problem or no though.
Gülşah Torkay I agree with Jorgaq's note that the caps for these tubes should be red, not blue. The similar blue cap for thickwall tubes will have a narrower stem that will not properly support the tube.
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