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YEPEdit

Yeast extract is what is sounds like, the extract of yeast thus it contains many nutrients yeast would normally use to grow

Bacto Peptone - is an enzymatic digest of animal protein (is a source of amino acids and of nitrogen, at least partially via amino acids e.g., glutamate, glutamine, proline). supposedly contains some trace glucose according to lab lore

We use Bacto peptone and Bacto yeast extract we have issues with other brands, so only use these to be safe...

For 1 L YEP (after autoclave and then adding sugar, it will be YEPD aka YPD, or YPG, etc)

10g Bacto Yeast Extract (i.e., final conc 1%)

20g Bacto Peptone (i.e., final conc 2%)

20g Agar (if making plates) (i.e., final conc 2%)

add water ~960ml if using glucose

autoclave in 2L flask

after cooling somewhat (~won't burn you)

add 40ml of 50% glucose (final concentration 2%) or 60ml 50% glycerol for 3%gly


FOR SD droupout media (1L) (e.g. -LEU, -etc)

1.7g YNB

5.0 g ammonium sulfate

X g of amino acid dropout mix (e.g., CSM-LEU) - the amount will be indicated on the bottle

water to 1L (minus the volume of sugar solution you will add after autoclaving)

autoclave, add sugar after autoclaving

if you are making SD media with agar, make it to 500ml (2X) and after autoclaving mix with 500ml 4%(2X) agar


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Synthetic media terminology and composition Edit

SD is synthetic defined media = YNB, Nitrogen, Carbon source, and CSM amino acids

SC complete media = YNB, Nitrogen, Carbon source, and SC amino acids.

YNB is yeast nitrogen base. Contains salts, vitamins and trace elements. Many companies YNB are lacking ammonium sulfate. Verify by the bottle or with the company whether product actually contains nitrogen. (details below)

DOB is dropout base = YNB + Nitrogen + Carbon source

Nitrogen = ammonium sulfate

Carbon source= usually the sugar glucose, or another carbon source, galactose, glycerol, ethanol, etc.

all of these can be +/- agar (sometimes they'll add an 'A' at the end of the name if containing agar. e.g., DOBA)

info from www.sunrisescience.com: and more info from MP; SC and CSM composition below are identical for sunrise and MP biosciences

Component in mg/L SC___ CSM___ Sandra___ Murakami et al Buffering paper
Adenine 21 10 19 40
L-Alanine 85.6 0 0 0
L-Arginine 85.6 50 19 20
L-Asparagine 85.6 0 0 0
L-Aspartic Acid 85.6 80 96 100
L-Cysteine 85.6 0 0 0
Glutamine 85.6 0 0 0
L-Glutamic Acid 85.6 0 96 100
Glycine 85.6 0 0 0
L-Histidine 85.6 20 19 100
Inositol 85.6 0 0 0
L-Isoleucine 85.6 50 77 0
L-Leucine 173.4 100 77 300
L-Lysine 85.6 50 58 30*
L-Methionine 85.6 20 19 20*
ParaAminoBenzoic Acid PABA

(also in YNB)

8.6 0 0 0
L-Phenylalenine 85.6 50 48 50
L-Proline 85.6 0 0 0
L-Serine 85.6 0 384 375
L-Threonine 85.6 100 192 200
L-Tryptophan 85.6 50 77 40
L-Tyrosine 85.6 50 58 30
Uracil 85.6 20 19 150
L-Valine 85.6 140 144 150

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for an unknown reason, Sandra has stated 'yeast don't like DI water' from the DI tap, so she uses 'glass-distilled water' instead. Sandra is the UW media person located in Foege

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Sunrise YNB mg/L____ MW (g/mol)____ µM_____ function
Potassium phosphate 1000
Inositol 2 180.16 11 structural basis for a number of secondary messengers
Copper sulfate (^assumed Cu(II) anyhydrous) 0.04 159.6 0.25^ cofactor
Magnesium sulfate 500 120.37 4154 cofactor
Niacin 0.4 123.1 3.25 precurser NAD/P, cofactor GPCR
Potassium iodide 0.1 166 0.60
Sodium chloride 100 58.44 1711
PABA 0.2 137.14 1.46 intermediate in the synthesis of folate, needed for DNA synth/repair, cofactor
Ferric chloride 0.2 162.2 1.23 cofactor, Fe/S clusters
Calcium chloride 100 110.98 901 cofactor
Pyridoxine 0.4 169.18 2.4  cofactor in many enzyme reactions in amino acid, glucose, and lipid metabolism
Manganese sulfate 0.4 151 2.6 cofactor
Biotin 0.002 244.3 0.01 a coenzyme for carboxylase enzymes, involved in the synthesis of fatty acids, isoleucine, and valine, and in gluconeogenesis.
Riboflavin 0.2 376.36 0.53 component of the cofactors FAD and FMN and as such required for a variety of flavoprotein enzyme reactions including activation of other vitamins
Sodium molybdate 0.2 205.92 0.97 cofactor (oxidation (hydroxylation) of xanthine and other purines and aldehydes. )
Pantothenate, calcium 0.4 238.27 1.7 to synthesize coenzyme-A (CoA), as well as to synthesize and metabolize proteins, carbohydrates, and fats.
Thiamine 0.4 300.8 1.3 derivative TPP - a coenzyme in the catabolism of sugars and amino acids
Zinc sulfate 0.4 161.47 2.5 cofactor
Folic acid 0.002 441.4 0.005 needed for DNA synth/repair, cofactor
Boric acid 0.5 61.83 8.1  promote amidations of carboxylic acids and amines in catalytic amounts?
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