Hi folks.
So my lab just got a refurbished 70TI rotor for ultracentrifuging but the product that arrived is much more banged up than the web advertised. I looked up the serial number and it is 15 years old (retirement age for this rotor is 12 years). It passed a test run up to speed and my co workers who have done other ultracentrifuge work seem to think I’m being a nervous nelly for being concerned about using this thing. I was trained very strictly that the metal would stress after a decade or so and could snap causing massive damage to the centrifuge. I need this part to do my lab work well but also know we cannot really afford a new rotor.
Any advice? I don’t want to be responsible for an 100,000 g force explosion.
Update: images of the ultracentrifuge explosion helped and we are working on getting a rotor we can know is safe! Thanks folks!
Our floor has an old Beckman optima-XL-100K ultracentrifuge. We keep getting the error referring to an unknown or bad overspeed disk. We replaced the overspeed disk with the correct one but this did not solve the problem. It gets up to 10-20K and then we get the unknown/bad disk error. Does anyone have any other suggestions about any other ways to solve this. It's my understanding that beckman has already refused to service this product due to it's age. At least that's my understanding based off of others who have tried in the past.
Hi everyone,
My friend is making lentivirus for the first time, but her lab does not have an ultracentrifuge to concentrate the virus. We have tried searching online but basically everything we found has told us to use the ultracentrifuge. Does anyone have any suggestions on what she could do? Any help would be appreciated! Thank you. :)
Hello,
I work at a biotech company and am currently part of a small group working to get an analytical ultracentrifuge method up and running (on top of the many, many other responsibilities our larger team as a whole handles). For those of you without a biology/biophysics background, an analytical ultracentrifuge measures absorbance, fluorescence, etc of material (usually protein, sometimes viruses or other biological particles) as it spins at high RPM and sediments to the outside of a cell over time. From this, we can determine things like how dense the particles in our sample are, which in my case is important because we want to know about how many virus particles are empty versus full.
In learning how to analyze these files, I realized that it's super complicated and could be made easier with a bit of automation to organize them. Basically, you start out with a .tar.gz file, which you then unzip to a .tar, which you unzip again to get a directory with a bunch of files. These files are named .Rxy, where x = A or I (for absorbance or intensity) and y = an integer between 1 and 8 inclusive (to note the cell number)- so for example, .RA2 is an absorbance scan for cell 2, or .RI6 is an intensity scan for cell 6. I have made a simple batch file to create a folder structure for this, which already helps a bunch, but I want to know if it's possible to move files into specific folders based on their extension. The batch file is below:
@echo off
mkdir AUC
mkdir AUC\cell1
mkdir AUC\cell1\absorbance
mkdir AUC\cell1\intensity
mkdir AUC\cell1\sunid(L)
mkdir AUC\cell1\runid(R)
mkdir AUC\cell2
mkdir AUC\cell2\absorbance
mkdir AUC\cell2\intensity
mkdir AUC\cell2\sunid(L)
mkdir AUC\cell2\runid(R)
mkdir AUC\cell3
mkdir AUC\cell3\absorbance
mkdir AUC\cell3\intensity
mkdir AUC\cell3\sunid(L)
mkdir AUC\cell3\runid(R)
mkdir AUC\cell4
mkdir AUC\cell4\absorbance
mkdir AUC\cell4\intensity
mkdir AUC\cell4\sunid(L)
mkdir AUC\cell4\runid(R)
mkdir AUC\cell5
mkdir AUC\cell5\absorbance
mkdir AUC\cell5\intensity
mkdir AUC\cell5\sunid(L)
mkdir AUC\cell5\runid(R)
mkdir AUC\cell6
mkdir AUC\cell6\absorbance
mkdir AUC\cell6\intensity
mkdir AUC\cell6\sunid(L)
mkdir AUC\cell6\runid(R)
mkdir AUC\cell7
mkdir AUC\cell7\absorbance
mkdir AUC\cell7\intensity
mkdir AUC\cell7\sunid(L)
mkdir AUC\cell7\runid(R)
mkdir AUC\cell8
mkdir AUC\cell8\absWe had an issue recently and I'm trying to figure out if it's normal, or if it's not, whether it's a user or equipment failure. Thanks!
Hello, i want to use ultracentrifugation to separate viruses fraction from gastric samples. We want to separate them by syringe and needle but have a problem with buying specialised tubes. I was looking by local distributors but no even one could help us. Any ideas which phrases I could search by to find them? I used to ask for thin walled test tubes for CsCl centrufiguation. Maybe you know any global distributors who can provide them? Thanks in advance!
We have a myriad of barely-functional ultracentrifuges in my building, so we constantly have to switch rotors out between whatever machine is working for the day. Today, somehow my list of ultracentrifuge malfunctions has increased to include a stuck rotor!
We are using the Beckman Coulter Avanti J series, and my JA-17 rotor won't come off of the pin no matter how hard I tug at it. I'm baffled. Has this ever happened to anyone else? And if so, HOW DO YOU GET THE DAMN THING OFF?
Signed, A sad maxiprep.
Mondays are so boring. This helped me pass a few minutes.
I’m new to UC for separation in a gene therapy process. I am wondering if anyone has any go to literature for UC principles/theory.
Decided to check a lab-mates spin before I left and it had stopped due to an error within the last hour. Now it doesn't want to start and I might have to lug a 25 pound rotor with samples across campus.
We really need a new centrifuge.
I have a question related to interpreting data obtained from Analytical Ultracentrifugation (AUC) (data below).
Sedimentation Equilibrium experiment
So for AUC which I am entirely unfamiliar with, how do you get MW from this type of plot? I understand how the OD will relate to it but I'm not clear on how the distribution of absorbance over the cell position yields a molecular weight. Any resources to help would be appreciated - I am not looking to perform this experiment as I do computational research, I am just trying to understand this plot from a paper I'm presenting on.
Hey! I’ve been trying to transduce this difficult cell line for some time now, and I figure that maybe concentrating viral titer will help with my experiment. I’m planning on doing some virus ultracentrifugation later today and I wanted to ask if any of you had found an optimal RPM for this protocol.I’ve seen numbers ranging from 16,000G through 25,000G for 2 hours. For reference, I’ll be using Beckman Ultra Clear tubes in an SW28 swinging bucket rotor.
Also, there seems to be a lack of consensus on what to do with regards to viral pellet resuspension. Some say to resuspend immediately, others suggest drying out at 4 degrees overnight, and I’ve seen a protocol where they add resuspension media on top of the pellet and leave it at 4 degrees overnight so it dissolves. Any thoughts on which approach might be best?
Thanks!
So I just ultracentrifuged some solubilized protein that I had extracted from biomass earlier today, and I realized that I hadn’t put the rotor back in the fridge, so the whole time, the ultracentrifuge was at around 13C, when we typically handle everything around 4C (the protein purification and solubilization was mostly done at room temperature, though ice was used for some steps). Is it possible that my protein has been ruined? It was only for maybe 30 minutes, and I immediately put it in the cold room afterwards. Not sure if I’m giving enough info though.
Phil
Sudden Lee
Go post NSFW jokes somewhere else. If I can't tell my kids this joke, then it is not a DAD JOKE.
If you feel it's appropriate to share NSFW jokes with your kids, that's on you. But a real, true dad joke should work for anyone's kid.
Mods... If you exist... Please, stop this madness. Rule #6 should simply not allow NSFW or (wtf) NSFL tags. Also, remember that MINORS browse this subreddit too? Why put that in rule #6, then allow NSFW???
Please consider changing rule #6. I love this sub, but the recent influx of NSFW tagged posts that get all the upvotes, just seem wrong when there are good solid DAD jokes being overlooked because of them.
Thank you,
A Dad.
So far nobody has given me a straight answer
Because a toothbrush works better
