I prepared the stock solution of 10X YNB (dissolve 134 g of yeast nitrogen base in 1L water and filtrate) for BMGY and BMMY preparation (media recipe from invitrogen) , I got the preblems
1. During I dissolved 10X YNB, I heated the solution for easy dissolving but it did not dissolved completely. How can I solve this problem?
2. I can filtrate it with 0.45 micron but not with 0.22 micron
3. I saw invitrogen protocol, it recommends that
"Dissolve 134 g of YNB with ammonium sulfate and w/o amino acids in 1L water" or " alternatively, use 34 g of YNB w/o ammonium sulfate and amino acids and 100 g of ammonium sulfate"
- I suspected that the amount of YNB in 1L is different between 2 preparing methods, doesn't it?
It doesn't affect to yeast growth, does it?
If it affect, what is the better one for Pichia culture?
Thank you
Hi there,
Actually it is the YNB which is not the same in the 2 recipes. In the first one the powder is containing ammonium sulfate, for the second one there is no ammonium sulfate in it (so it is the one to consider if you want to provide the yeast with an other source of nitrogen). So the final concentration of ammonium sulfate is 10% for both recipes but you have to make sure you are using the right powder!
YNB is indeed not fully soluble (I'm using it at 67g/L as a 10x stock for cerevisiae and there is always a cloudy aspect before filter sterilization). But filtration on 0.22µm is definitely not a problem (do it under vacuum if needed). What you can also try for improving dissolution of the powder is sonication...
Hi Thatcheewa
You use 1.7 g/L of YNB (w/o ammonium sulfate and amino acids) in Yeast Minimal Media/Synthetic Defined (SD) for Saccharomyces cerevisiae .
Good Luck !!!
I didn't realize and have filtered plus autoclaved the YNB w ammonium sulfate media (with added dextrose anhydrous) -----how will this affect the media composition; is it really bad; I didn't realize and have grown in previously only autoclaved media and they did grow but I am curious as to how heat may be detrimental to this media; is it okay to autoclave a media with YNB(w ammonium sulfate) and glucose? Thanks!!
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