McClean:KS Amino Acid Supplement

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Summary

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.

Mixing Tips

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
uracil MP Biomedicals Uracil 103204 20 200
tryptophan Fisher L-tryptophan, tissue culture grade BP395 50 500
histidine-HCl Fisher L-histiine, mono HCL monohydrate BP383 20 200
arginine-HCl Fisher L-arginine HCL BP372 20 200
methionine Fisher L-methionine BP388 20 200
tyrosine Fisher L-tyrosine BP396 30 300
leucine Fisher L-leucine BP385 100 1000
isoleucine Fisher L-isoleucine BP384 30 300
lysine-HCl Fisher L-lysine monohydrochloride BP386 100 1000
phenylalanine Fisher L-phenylalanine BP391 50 500
glutamic acid Fisher L-glutamic acid monosodium salt 194677 100 1000
aspartic acid Fisher L-aspartic acid BP374 100 1000
valine Fisher valine BP397 150 1500
threonine Fisher L-threonine BP394 200 2000
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
uracil MP Biomedicals Uracil 103204 2000 2 1
tryptophan Fisher L-tryptophan, tissue culture grade BP395 5000 5 2.5
histidine-HCl Fisher L-histiine, mono HCL monohydrate BP383 2000 2 1
arginine-HCl Fisher L-arginine HCL BP372 2000 2 1
methionine Fisher L-methionine BP388 2000 2 1
tyrosine Fisher L-tyrosine BP396 3000 3 1.5
leucine Fisher L-leucine BP385 10000 10 5
isoleucine Fisher L-isoleucine BP384 3000 3 1.5
lysine-HCl Fisher L-lysine monohydrochloride BP386 10000 10 5
phenylalanine Fisher L-phenylalanine BP391 5000 5 2.5
glutamic acid Fisher L-glutamic acid monosodium salt 194677 10000 10 5
aspartic acid Fisher L-aspartic acid BP374 10000 10 5
valine Fisher valine BP397 15000 15 7.5
threonine Fisher L-threonine BP394 20000 20 10
serine Acros Organics L-serine 99% 132661000 40000 40 20

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

  1. List troubleshooting tips here.
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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.

References

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.

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