Media Library

* Add 5% defibernated sheep blood using sterile technique. Allow blood to come to room tempurature before adding to media.
* Below is recipe for Blood Agar
	+ 10.0 g Tryptose
	+ 3.0 g Beef extract
	+ 5.0 g sodium chloride
	+ 15.0 g agar
	+ 1000mL media water
	+ final pH 7.0 +- 0.2

### Borate Buffering Agar???

* 15mL Borate Buffer
* 3g iron ion agar
* 273mL saline
* 3mL Merthiolate solution (1 1/2 in saline, mix and heat, do not sterilize, dispense in 4mL tubes)

### Brilliant Green Lactose Bile Broth

* M10g Peptone
* 10g Lactose
* 20g Oxgall
* 0.0133g Brilliant Green
* 1000mL DI H2O

### CaCl2 (50mM pH 6.1)

* 7.3505 g CaCl2 dihydride
* 1000mL DI H2O
* autoclave

### Casein Agar

* 2mL sterile skimmed milk (in petri dish)
* plus Casein agar base
* 2g peptone
* 1g glucose
* 2g Na2HPO4
* 10g NaCl
* 15g Agar
* 1000mL DI H20

### Citrate Medium (Koser's)

* 1.5g NaNH4HPO4
* 1.0g KH2PO4
* 0.2g MgSO4
* 3.0g Sodium Citrate
* 1000mL DI H2O

### Citrate Agar (Simmon's)

* 0.2g MgSO4
* 1.0g NH4H2PO4
* 1.0g K2HPO4
* 2.0g Sodium Citrate
* 5.0g NaCl
* 15.0g Agar
* 0.08g Brom Thymol Blue
* 1000mL DI H2O

### Desoxycholate Agar

* 10g Peptone
* 10g Lactose
* 1.0g Sodium Desoxycholate
* 5g Sodium Chloride
* 2g Dipotassium phosphate
* 1g Ferric Citrate
* 1g Sodium Citrate
* 15g Agar
* 0.03g Neutral Red
* 1000mL DI H2O

### Desulfovibrio Medium

* Makes 1 Liter standard mineral base, pH 7.2
* 1.0g NH4Cl
* 5.0g Na2SO4
* 4.0mL 95% EtOH
* 1000mL DI H2O

### Dextrose-Peptone-Methyl Red Broth

* 1000mL DI H2O
* 5.0g Proteose Peptone
* 5.0g Dextrose
* 5.0g Dipotassium Phosphate K2HPO4
* Adjust pH to 6.9

### Eosin Methylene Blue Agar (EMB Agar)

* 10g Peptone
* 10.0g Lactose
* 5.0g Saccharose
* 2.0g Dipotassium Phosphate (K2HPO4)
* 15.0g Agar
* 0.4g Eosin Y
* 0.065g Methylene Blue
* 1000mL DI H2O
* adjust to pH 7.2

### Endo Agar

* 10.0g Peptone
* 10.0g Lactose
* 3.5g Dipotassium Phosphate
* 15g Agar
* 2.5g Sodium Sulfite
* 0.4g Basic Fuchsin
* 1000mL DI H2O

### Glucose Yeast Extract

* 10g Tryptone
* 10g Yeast Extract
* 5g Dipotassium Phosphate (K2HPO4)
* 5g Glucose
* 15g Agar
* 1000mL DI H2O

### Glycerol Yeast Extract Agar

* 5.0g Yeast Extract
* 50.0mL Glycerol
* 15.0g Agar
* 1000mL DI H2O

### Koser's Citrate Medium

* 1.5g Sodium ammonium phosphate
* 1.0g Dipotassium Phosphate (K2HPO4)
* 0.2g MgSO4
* 3.0g Sodium citrate
* 1000mL DI H2O
* Adjust to pH 6.7

### **LB Agar**

* To make 1 Liter, follow the recipe for LB Broth Medium and then add 15g of agar, mix on heat until in solution and then autoclave.

### **LB + amp agar**

* After autoclaving LB agar, cool to 50°C (no higher) in a temperature-controlled water bath.
* 0.6mL **Ampicillin** solution.  (**100mg ampicillin/mL Stock > 1 mL Stock / 1 L LB agar**)
	+ Or 100ug ampi/mL agar

Look at this generalized recipe for our new ampicillin: HERE

or here

### **Luria-Bertani Broth Medium(LB)**

* Makes 1 Liter

Adjust the pH to 7.0 - 7.4 with 5 M NaOH (~0.2 mL).
Adjust the final volume of the solution to 1 L with H2O.
Sterilize by autoclaving for 20 min at 15 psi (1.05 kg/cm2) on liquid cycle.

### M.R. - V.P. Medium

* Makes 1 Liter

 Dextrose
 Dipotassium Phosphate

* Methyl Red solution in dropper: Prepare the methyl red indicator by dissolving 0.1 g of methyl

red in 300 mL of 95% ethyl alcohol. Add sufficient purified
water to make 500 mL.

* Voges-Proskauer solution in dropper: Berrit's Reagent A and Reagent B

### Nitrate Broth

* Have premade media + durham tube = 6mL in 16x125tt

### **Penassay Agar**

* Makes 1 Liter

 Bacto-Beef Extract
 Bacto-Yeast Extract
 Bacto-Peptone
 Glucose
 Sodium Chloride NaCl
 Dipotassium Phosphate
 Monopotassium Phosphate

### **Tryptic Soy Agar plus Lactose (TSA + )**

Instructions with TSB

 5.0g
 5.0g
 0.2g
 18.0g
 1.0L

Instructions with TSA

 5.0 g
 5.0 g
 0.2 g
 1.0 L

### **Urea Broth**

 9.1 g/L
 0.01 g/L
 0.1 g/L
Filter sterilize : DO NOT HEAT
When the powder is dissolved, sterilized by filtration.
Dispense quantities of 0.5 to 2 ml in small sterile tubes. Larger volumes can be used but the reactions will be slower.
Do not sterilize in autoclave.
Do not boil the medium. When there is no filter available, the medium can be sterilized at 100 -110°C for 10 minutes. If the medium is prepared and inoculated immediately it provides good results without sterilizing. The prepared medium should be stored at 2-8°C. The color is red-orange.

### **Yeast Extract Peptone Dextrose YPD (YEPD) Medium (Broth or Agar)**

* Omit agar for liquid culture.
* Agar plates are at 2% rather than the 1.5% usual in bacterial work.
* Dry the plates 2-3 days at RT before storing at 4C for several months.
* Autoclave components for a maximun of 15 minutes at 121C.
* For large volumes, autoclave salts, glucose and agar separately.

A complex medium for routine growth. Available premixed as Difco 0428-17.

* per liter
* Bacto yeast extract (Difco 0127-17) (1%) 10g
* Bacto peptone (Difco 0118-17) (2%) 20g
* Glucose (2%) 20g
* Bacto agar (Difco 0140-01) (2%) 20g