McClean:KS Amino Acid Supplement
This is the amino acid supplement used in our low fluorescence media and synthetic complete medias. To make dropout media, leave out the appropriate amino acid (ie, to make SC-URA, leave out uracil). These supplements are stored as dry powder stocks on the media shelf in brown bottles (some amino acids are light sensitive). If you need to remake a supplement stock, please use exactly the amino acids listed (take note of catalog numbers) and clearly initial and date the bottle.
Do not autoclave amino acids, as they may change their chemical structure. To mix them into a liquid solution, first microwave the liquid solution until it is warm but not painfully hot to the touch. Then, slowly add in the dry powder while the solution is stirred with a magnetic stir bar. It is the clumps which dissolve slowly. If this is done correctly, a solution can be prepared in 20 min. If they dry powder is poured in in clumps or if the water is initially cold, it will take hours to dissolve.
Final Composition in Liquid Media
|Amino Acid||Supplier||Description||Catalog No.||KS Supp Final (mg/L)||KS Supp 10x stock (mg/L)|
|adenine hemisulfate||Sigma||Adnine hemisulfate salt, min 99%||A9126||50||500|
|tryptophan||Fisher||L-tryptophan, tissue culture grade||BP395||50||500|
|histidine-HCl||Fisher||L-histiine, mono HCL monohydrate||BP383||20||200|
|glutamic acid||Fisher||L-glutamic acid monosodium salt||194677||100||1000|
|aspartic acid||Fisher||L-aspartic acid||BP374||100||1000|
|serine||Acros Organics||L-serine 99%||132661000||400||4000|
Dry Powder Stocks
To make dry stocks mix all items together (leave out specific amino acids if desired) in a brown plastic bottle, turn end over end MANY MANY times to mix with a sterile marble added to aid mixing. Sterile marbles are stored on the media shelf. Store the final mix dry on the media shelf in a brown opaque bottle. Initial and clearly date your new amino acid stock.
|Amino Acid||Supplier||Description||Catalog No.||Dry power enough for 10-1L 10x stocks (mg)||Dry power enough for 10-1L 10x stocks (grams)||Dry power enough for 5-1L 10x stocks (grams)|
|adenine hemisulfate||Sigma||Adnine hemisulfate salt, min 99%||A9126||5000||5||2.5|
|tryptophan||Fisher||L-tryptophan, tissue culture grade||BP395||5000||5||2.5|
|histidine-HCl||Fisher||L-histiine, mono HCL monohydrate||BP383||2000||2||1|
|glutamic acid||Fisher||L-glutamic acid monosodium salt||194677||10000||10||5|
|aspartic acid||Fisher||L-aspartic acid||BP374||10000||10||5|
|serine||Acros Organics||L-serine 99%||132661000||40000||40||20|
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- List troubleshooting tips here.
- You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
- Anecdotal observations that might be of use to others can also be posted here.
Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.
- Taylor D. Scott 17:49, 11 July 2017 (EDT): L-Glutamic acid monosodium salt hydrate from Sigma (catalog no. G1626) has the same CAS number and FW as the Fisher L-Glutamic acid monosodium salt (catalog no. 194677). I used the same amount (10g) when making the latest KS supplement dry powder stock.
This amino acid mix was developed by Katja Schwartz (KS) in the Botstein lab at Stanford University. It is adapted from the synthetic complete dropout mix in the CSH Yeast Genetics Course lab manual.
- Megan N McClean 14:01, 25 August 2011 (EDT)
or instead, discuss this protocol.