McClean:KS Amino Acid Supplement

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.

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!

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References

This amino acid mix was developed by Katja Schwartz in the Botstein lab at Stanford University. It is adapted from the synthetic complete dropout mix in the CSH Yeast Genetics Course lab manual.

Contact

• Megan N McClean 14:01, 25 August 2011 (EDT)

or instead, discuss this protocol.