Hello, I prepared TB media for BL21 protein expression but didn't include glycerol, will lack of this carbon source affect the quality of protein expressed?
Dear Maya Kornaj ,
I presume you used the following protocol:
http://cshprotocols.cshlp.org/content/2015/7/pdb.rec087874.full?text_only=true
(in your case without glycerol…). According to: https://khimexpert.com/wp-content/uploads/2018/12/GCM07-Terrific-Broth.pdf it states “Tryptone and Yeast Extract are the sources for carbon, nitrogen, vitamins, minerals, and amino acids essential for growth, whereas the potassium phosphate buffer system acts to prevent cell death.” And “Glycerol serves an additional source for carbon as well as a source for carbohydrates ...”
So, you can give it a go but being on the save side you better make new TB media.
Best regards.
Not necessarily the quality, but certainly the quantity. Last time I forgot to add it, I ended up with fewer cells making less protein by about a factor of 5.
Maya Kornaj Using TB media without glycerol may lead to cultures that alkalize to pH 8.3 overnight, that is higher than in standard TB media having glycerol which keeps the pH 7.0 during overnight incubation. The presence of glycerol appears to significantly delay alkalization in standard TB media. Thus TB (-Glycerol)cultures may lead to smaller yield/smaller pellet.
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