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Y 1 Adrenal Cell Growth Medium
Composition per 101.0mL:
Ham’s F-10 medium ................................... .90.0mL
Fetal bovine serum....................................... .10.0mL
Penicillin-streptomycin solution................... ...1.0mL
pH 7.0 ± 0.2 at 25oC
Ham’s F-10 Medium:
Composition per liter:
NaCl.................................................................. ..7.4g
NaHCO3........................................................... ..1.2g
Glucose ........................................................... ...1.1g
NaH2PO4*H2O............................................... ...0.29g
KCl.................................................................. ..0.28g
L-Arginine*HCl............................................... ..0.21g
L-Glutamine ................................................... ...0.15g
MgSO4*7H2O.................................................. .0.15g
Sodium pyruvate............................................. ..0.11g
KH2PO4........................................................... .0.08g
CaCl2*2H2O .................................................. ...0.04g
L-Cystine*2HCl ............................................... .0.04g
L-Histidine*HCl*H2O...................................... ..0.02g
L-Lysine*HCl.................................................. ..0.02g
L-Asparagine-H2O......................................... ...0.01g
L-Aspartic acid................................................ ..0.01g
L-Glutamic acid................................................ .0.01g
L-Leucine ......................................................... .0.01g
L-Proline......................................................... ...0.01g
L-Serine............................................................ .0.01g
L-Alanine......................................................... .8.9mg
Glycine............................................................ .7.5mg
D-Phenylalanine ............................................. ..5.0mg
L-Methionine................................................... .4.5mg
Hypoxanthine................................................ ...4.1mg
L-Threonine................................................... ...3.6mg
L-Valine......................................................... .. 3.5mg
L-Isoleucine................................................... ...2.6mg
L-Tyrosine ..................................................... ..1.8mg
Vitamin B12................................................... ...1.4mg
Folic acid ...................................................... ...1.3mg
Phenol Red...................................................... .1.2mg
Thiamine*HCl ............................................... ..1.0mg
FeSO4*7H2O ................................................. .0.8mg
Choline chloride............................................. ..0.7mg
D-Calcium pantothenate.................................. ..0.7mg
Thymidine...................................................... ..0.7mg
Niacinamide................................................... ..0.6mg
L-Tryptophan................................................... .0.6mg
Isoinositol ..................................................... ..0.5mg
Riboflavin ..................................................... ..0.4mg
Lipoic acid ...................................................... .0.2mg
Pyridoxine*HCl............................................. ...0.2mg
ZnSO4*7H2O............................................... ..0.03mg
Biotin ........................................................... .0.02mg
CuSO4*5H2O.................................................. ..3.0ตg
Preparation of Ham’s F-10 Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Penicillin-Streptomycin Solution:
Composition per 100.0mL:
Streptomycin...................................................... .0.5g
Penicillin G ............................................ ...500,000U
Preparation of Penicillin-Streptomycin Solu-
tion: Add components to distilled/deionized water
and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.
Preparation of Medium: Aseptically combine components. Filter sterilize. Store at 4-5oC.
Use: For the cultivation of Y-1 mouse adrenal tissue culture cells used for the detection of heat-labile toxin (LT) produced by enterotoxigenic strains of Escherichia coli. LT causes the conversion of elongated fibroblast-
like cells into round, refractile cells.
Y 1 Adrenal Cell Growth Medium
Composition per 580.0mL:
Ham’s F-10 medium ................................ ..500.0mL
Fetal bovine serum....................................... .75.0mL
Penicillin-streptomycin solution................... ...5.0mL
pH 7.0 ± 0.2 at 25oC
Ham’s F-10 Medium:
Composition per liter:
NaCl................................................................. ..7.4g
NaHCO3........................................................... ..1.2g
Glucose ........................................................... ...1.1g
NaH2PO4*H2O.............................................. ...0.29g
KCl................................................................. ..0.28g
L-Arginine*HCl............................................... ..0.21g
L-Glutamine..................................................... .0.15g
MgSO4*7H2O.................................................. .0.15g
Sodium pyruvate............................................. ..0.11g
KH2PO4........................................................... .0.08g
CaCl2*2H2O..................................................... .0.04g
L-Cystine*2HCl............................................... ..0.04g
L-Histidine*HCl*H2O...................................... ..0.02g
L-Lysine*HCl.................................................. ..0.02g
L-Asparagine-H2O......................................... ...0.01g
L-Aspartic acid ............................................... ..0.01g
L-Glutamic acid................................................ .0.01g
L-Leucine........................................................ ..0.01g
L-Proline........................................................ ...0.01g
L-Serine ........................................................... .0.01g
L-Alanine........................................................ .8.9mg
Glycine........................................................... .7.5mg
D-Phenylalanine ............................................ ..5.0mg
L-Methionine................................................... .4.5mg
Hypoxanthine............................................... ...4.1mg
L-Threonine................................................... ...3.6mg
L-Valine ....................................................... .. 3.5mg
L-Isoleucine................................................... ...2.6mg
L-Tyrosine..................................................... ...1.8mg
Vitamin B12................................................... ...1.4mg
Folic acid ...................................................... ...1.3mg
Phenol Red...................................................... .1.2mg
Thiamine*HCl ............................................... ..1.0mg
FeSO4*7H2O ................................................. .0.8mg
Choline chloride............................................. ..0.7mg
D-Calcium pantothenate.................................. ..0.7mg
Thymidine...................................................... ..0.7mg
Niacinamide................................................... ..0.6mg
L-Tryptophan................................................... .0.6mg
Isoinositol ..................................................... ..0.5mg
Riboflavin ..................................................... ..0.4mg
Lipoic acid ...................................................... .0.2mg
Pyridoxine*HCl............................................. ...0.2mg
ZnSO4*7H2O............................................... ..0.03mg
Biotin ........................................................... .0.02mg
CuSO4*5H2O.................................................. ..3.0ตg
Preparation of Ham’s F-10 Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly.
Penicillin-Streptomycin Solution: Composition per 100.0mL:
Streptomycin...................................................... .0.5g
Penicillin G............................................. ...500,000U
Preparation of Penicillin-Streptomycin Solu-
tion: Add components to distilled/deionized water
and bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.
Preparation of Medium: Aseptically combine
components. Filter sterilize. Store at 4o-5oC.
Use: For the cultivation of Y-1 mouse adrenal tissue cul-
ture cells used for the detection of cholera enterotoxin
(CT) produced by enterotoxigenic strains of Vibrio chol-
erae or Vibrio mimicus. CT causes the conversion of
elongated fibroblast-like cells into round, refractile cells.
Yeast Extract Agar
Composition per liter:
Agar .............................................................. ...15.0g
Peptone ........................................................... ...9.5g
Yeast extract...................................................... ..7.0g
Beef extract...................................................... ...5.0g
NaCl.................................................................. ..5.0g
pH 7.0 ± 0.2 at 25oC
Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure-121oC. Pour into sterile Petri dish-
es or leave in tubes.
Use: For the cultivation of Aeromonas salmonicida.
Yeast Extract Agar
Composition per liter:
Agar .............................................................. ...20.0g
Yeast extract..................................................... ..1.0g
Buffer solution ............................................ ...2.0mL
pH 6.0 ± 0.2 at 25oC
Buffer Solution:
Composition per 400.0mL:
KH2PO4........................................................... .60.0g
Na2HPO4........................................................ ..40.0g
Preparation of Buffer Solution: Add 40.0g of Na2HPO4 to 300.0mL of distilled/deionized water. Mix thoroughly. Add 60.0g of KH2PO4. Mix thor-
oughly. Adjust pH to 6.0.
Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Autoclave for 15 min at 15 psi pres-
sure-121oC. Pour into sterile Petri dishes.
Use: For the identification of Histoplasma capsulatum,
Blastomyces dermatitidis, and Coccidioides immitis.
Yeast Extract Agar
Composition per liter:
Agar .............................................................. ...15.0g
Proteose peptone............................................. ..10.0g
NaCl................................................................. ..5.0g
Yeast extract..................................................... ..3.0g
Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure-121oC. Pour into sterile Petri dish-
es or leave in tubes.
Use: For the cultivation of a variety of heterotrophic microorganisms.
Yeast Extract Sodium Lactate Medium
Composition per liter:
Agar .............................................................. ...15.0g
Pancreatic digest of casein.............................. ..10.0g
Yeast extract.................................................. ...10.0g
Sodium lactate................................................ ...10.0g
KH2PO4........................................................... ...2.5g
MnSO4 .......................................................... .5.0mg
pH 7.0 ± 0.2 at 25oC
Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure-121oC. Pour into sterile Petri dish-
es or leave in tubes.
Use: For the isolation, cultivation, and maintenance of Propionibacterium species.
Yeast Extract Sucrose Agar
(YESA)
(ATCC Medium 2125)
Composition per liter:
Sucrose........................................................... ..20.0g
Agar .............................................................. ...15.0g
Yeast extract...................................................... ..4.0g
KH2PO4........................................................... ...1.0g
MgSO4*7H2O.................................................. ...0.5g
pH 6.2 ± 0.2 at 25oC
Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 6.2. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto-
clave for 15 min at 15 psi pressure-121oC. Pour into sterile Petri dishes or leave in tubes.
Use: For the cultivation and maintenance of various
fungi.
Yeast Extract Sucrose Agar
Composition per liter:
Sucrose........................................................ ...150.0g
Agar .............................................................. ...20.0g
Yeast extract................................................... ...20.0g
MgSO4*7H2O.................................................. ...0.5g
Trace metal solution....................................... . 1.0mL
pH 6.5 ± 0.2 at 25oC
Trace Metal Solution:
Composition per 100.0mL:
ZnSO4*7H2O..................................................... .1.0g
CuSO4*5H2O..................................................... .0.5g
Preparation of Trace Metal Solution: Add components to 100.0mL distilled/deionized water. Mix thoroughly.
Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Adjust pH to 6.5. Gently heat and bring to boiling. Distribute into tubes or flasks. Auto-
clave for 15 min at 15 psi pressure-121oC. Pour into sterile Petri dishes or leave in tubes.
Use: For the cultivation and maintenance of various
fungi.
Yeast Fermentation Medium
Composition per liter:
Peptone ........................................................... ...7.5g
Yeast extract...................................................... ..4.5g
Bromthymol Blue (1.6% solution)............... ...1.0mL
Carbohydrate solution.................................... .1.0mL
Carbohydrate Solution:
Composition per 10.0mL:
Carbohydrate..................................................... .0.6g
Preparation of Carbohydrate Solution: Add
carbohydrate to distilled/deionized water and bring
volume to 10.0mL. Glucose, maltose, lactose, galac-
tose, or trehalose may be used. If raffinose is used, pre-
pare a 12% solution. Mix thoroughly. Filter sterilize.
Preparation of Medium: Add components, ex-
cept carbohydrate solution, to distilled/deionized wa-
ter and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute in 2.0mL vol-
umes into test tubes that contain an inverted Durham tube. Autoclave for 15 min at 15 psi pressure-121oC. Cool to 45o-50oC. Aseptically add 1.0mL of sterile carbohydrate solution. Mix thoroughly.
Use: For the cultivation and differentiation of yeast
based on carbohydrate fermentation patterns. Yeasts
that can ferment a specific carbohydrate turn the medi-
um yellow.
Yeast Glucose Agar
Composition per liter:
Agar .............................................................. ...15.0g
Pancreatic digest of gelatin............................... .7.75g
Beef extract...................................................... .4.75g
Yeast extract..................................................... ..2.5g
K2HPO4........................................................... ...2.5g
Glucose ........................................................... ...1.0g
pH 7.0 ± 0.2 at 25oC
Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure-121oC. Pour into sterile Petri dish-
es or leave in tubes.
Use: For the cultivation and maintenance of a wide va-
riety of bacteria.
Yeastrel Agar
Composition per liter:
Agar .............................................................. ...15.0g
Peptone.............................................................. .9.5g
Yeastrel.............................................................. .7.0g
Lab-lemco (meat extract).................................. ...5.0g
NaCl................................................................. ..5.0g
pH 7.0 ± 0.2 at 25oC
Source: Lab-lemco is available from Oxoid Uni-
path. Yeastrel is produced by Mapleton’s Foods Ltd.,
Moss Street, Liverpool and is available from health
food shops worldwide.
Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure-121oC. Pour into sterile Petri dish-
es or leave in tubes.
Use: For the cultivation of Aeromonas salmonicida.
YEPD Medium
Composition per liter:
Agar .............................................................. ...20.0g
Glucose ........................................................... .20.0g
Peptone ........................................................... .20.0g
Yeast extract................................................... ...10.0g
Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure-121oC. Pour into sterile Petri dish-
es or leave in tubes.
Use: For the cultivation of a variety of heterotrophic microorganisms.
Yersinia Selective Agar Base
Composition per liter:
Mannitol......................................................... ...20.0g
Peptone ........................................................... .17.0g
Agar .............................................................. ...12.5g
Proteose peptone................................................ .3.0g
Yeast extract...................................................... ..2.0g
Sodium pyruvate................................................ .2.0g
NaCl.................................................................. ..1.0g
Sodium desoxycholate....................................... .0.5g
MgSO4*7H2O.................................................. .0.01g
Neutral Red...................................................... .0.03g
Crystal Violet................................................ ...1.0mg
Selective supplement .................................... .. 6.0mL
pH 7.4 ± 0.2 at 25oC
Source: This medium is available as a premixed
powder from BD Diagnostic Systems and Oxoid Un-
ipath.
Selective Supplement:
Composition per 6.0mL:
Cefsulodin................................................... ...15.0mg
Irgasan........................................................... ..4.0mg
Novobiocin .................................................. ...2.5mg
Ethanol......................................................... ... 2.0mL
Preparation of Selective Supplement: Asepti-
cally add components to 4.0mL of distilled/deionized water and 2.0mL of ethanol. Mix thoroughly.
Preparation of Medium: Add components to distilled/deionized water and bring volume to 1.0L. Mix thoroughly. Gently heat and bring to boiling. Distribute into tubes or flasks. Autoclave for 15 min at 15 psi pressure-121oC. Cool to 50oC. Aseptically add selective supplement. Mix thoroughly. Pour into sterile Petri dishes or leave in tubes.
Use: For the isolation and enumeration of Yersinia en-
terocolitica from clinical specimens.
YI-S Medium
Composition per liter:
YI broth..................................................... .880.0mL
Bovine serum, heat inactivated................... .100.0mL
Vitamin mixture 18........................................ 20.0mL
Source: Vitamin mixture 18 is available from Bio-
fluids, Inc., Rockville, MD.
YI Broth:
Composition per liter:
YI base stock............................................... 780.0mL
10ื Glucose buffer stock........................... .100.0mL
YI Base Stock:
Composition per 780.0mL:
Yeast extract.................................................. ...30.0g
L-Cysteine*HCl ............................................... ...1.0g
NaCl................................................................. ..1.0g
Ascorbic acid .................................................. ...0.2g
Ferric ammonium citrate........................... ...228.0mg
10ื Glucose Buffer Stock:
Composition per 100.0.0mL:
Glucose ........................................................... .10.0g
K2HPO4........................................................... ...1.0g
KH2PO4........................................................... ...0.6g
Preparation of 10ื Glucose Buffer Stock:
Add components to distilled/deionized water and
bring volume to 100.0mL. Mix thoroughly. Filter
sterilize.
Preparation of YI Base Stock: Add components to 600.0mL of distilled/deionized water. Mix thor-
oughly. Bring volume to 780.0mL with distilled/ deionized water. Adjust pH to 6.8 with 1N NaOH. Distribute in 78.0mL aliquots to 100.0mL screw-
capped bottles. Autoclave for 15 min at 15 psi pres-
sure-121oC. Cool to room temperature.
Preparation of YI Broth: Aseptically add
10.0mL of 10ื glucose buffer stock to 78.0mL of
cooled YI base stock. Adjust osmolarity with NaCl to 380.0milliosmols/kg.
Preparation of Medium: Aseptically add 2.0mL of vitamin mixture 18 and 10.0mL of heat-inactivat-
ed bovine serum to 88.0mL of YI broth. Distribute in 13.0mL aliquots to 16 x 125mm screw-capped test tubes. Store at 4oC in the dark with the caps screwed on tightly. Use within 96h.
Use: For the cultivation of Entamoeba species.
YPC Medium
Composition per liter:
Agar .............................................................. ...15.0g
Proteose peptone............................................. ..15.0g
Yeast extract...................................................... ..5.0g
KH2PO4........................................................... ...4.0g
Sucrose.............................................................. .2.5g
Glucose ........................................................... ...2.0g
L-Cystine............................................................ .0.5g
Na2SO3.............................................................. .0.2g
pH 7.2 ± 0.2 at 25oC
Preparation of Medium: Add components to
distilled/deionized water and bring volume to 1.0L.
Mix thoroughly. Gently heat and bring to boiling.
Distribute into tubes or flasks. Autoclave for 15 min
at 15 psi pressure-121oC. Pour into sterile Petri dish-
es or leave in tubes.
Use: For the cultivation of Pasteurella multocida.
YPNC Medium
Composition per liter:
Agar .............................................................. ...18.0g
NaCl............................................................... ...2.92g
KH2PO4........................................................ ..0.596g
Yeast extract...................................................... ..0.5g
Sodium hydrogen glutamate (pH 6.0) ............ ..0.37g
K2HPO4........................................................ ..0.107g
NH4Cl ........................................................... .0.107g
MgSO4*7H2O............................................... ..0.049g
Glucose solution ......................................... . 10.0mL
Trace metals solution..................................... .. 1.0mL
pH 6.0 ± 0.2 at 25oC
Glucose Solution:
Composition per 10.0mL:
Glucose ........................................................... ...4.0g
Preparation of Glucose Solution: Add glucose to distilled/deionized water and bring volume to 10.0mL. Mix thoroughly. Filter sterilize.
Trace Metals Solution:
Composition per liter:
EDTA.............................................................. .50.0g
ZnSO4*7H2O ................................................. ..22.0g
CaCl2.............................................................. ..5.54g
MnCl2*4H2O ................................................. ..5.06g
FeSO4*7H2O................................................. ...4.99g
(NH4)6 Mo7O14*H2O ..................................... ..1.10g
CoSO4*5H2O.................................................. ..1.57g
CoCl2*6H2O ................................................. ...1.61g
Preparation of Trace Metals Solution: Add
components, one at a time, to distilled/deionized wa-
ter and bring volume to 1.0L. Mix thoroughly. Filter
sterilize.
Preparation of Medium: Add components, ex-
cept glucose solution and trace metals solution, to
distilled/deionized water and bring volume to
989.0mL. Mix thoroughly. Gently heat and bring to
boiling. Adjust pH to 6.0 with KOH. Autoclave for
15 min at 15 psi pressure-121oC. Cool to 50oC.
Aseptically add 10.0mL of sterile glucose solution
and 1.0mL of sterile trace metals solution. Mix thor-
oughly. Pour into sterile Petri dishes or aseptically
distribute into sterile tubes.
Use: For the cultivation and maintenance of a variety of Cryptococcus species.
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