I tried using ChatGPT for the first time...thought maybe it could provide something different than what I planned. I think it is giving really wrong information. Wouldn't using acetone precipitate macromolecules (and not small metabolites)?
I asked for a method to separate small metabolites from a cell lysis, ChatGPT replied with.....
Certainly! Here's a simple procedure that you could use for yeast lysis and crude purification of small metabolites:
I'm sure ChatGPT can be very convenient, and I've heard it can be used to develop protocols etc., but I'm not convinced.
Do not rely on ChatGPT to do your literature searching for you. It was not designed to be an authoritative source of information, and it is well-recognized that it often provides erroneous information. You would be better off using a search engine to find references, and then reading the references.
Acetone will precipitate proteins. Whether it precipitates small metabolites depends on the nature of the metabolites.
An easy way to remove macromolecules while retaining low molecular weight, soluble metabolites is to use an ultrafiltration device. Molecules larger than the molecular weight cutoff will be retained by the filter and small molecules will pass through.
You can concentrate the small-molecule filtrate by freeze-drying with a lyophilizer or Speed-Vac. Bear in mind that non-volatile salts will also be concentrated by freeze-drying.
Another way to dry down small-volume samples is to gently blow nitrogen gas over them while warming them in the hand, or using a device designed for this purpose.
Thank you Dr. Shapiro!
Yes, I have centrifugal filters the Amicon Ultra ones with a 10 kDal cut off. This is currently the best option I have.
I was just wondering whether there is a better known method for separating or filtering everything above ~0.5 kDal. Or if there is a convenient procedure for precipitating macromolecules.
I'm considering acetone or chloroform extraction, except I am worried that this would remove smaller metabolite in a variable way..
GPT is really interesting and I know it isn't correct. I'm interested in learning how to use it, if anything I think it would make a superior spelling/grammar checker.
Thank you again!
There probably is not one single method that will precipitate all macromolecules and no metabolites. Several different methods of separation may be needed. I suggest you read some metabolomics papers to find suitable methods.
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