LB (Luria-Bertani) medium


Commercially available: or use the following recipe:


To 950 mls of ultrapure water, add


10g bacto-tryptone


5g bacto-yeast extract


10g NaCl.


15g bacto-agar, to solidify.


Mix, autoclave. PH to 7 before adding agar.


Weigh agar (granulated agar) and place directly into final vessel –(agar will not dissolve until solution boils). Mix, autoclave. When cool enough (around 55 C), pour into dishes (in the fume hood) like 25ml/100mmm dishes.


LBamp: add 1 ml of the 100 mg/ml stock ampicillin to each liter of cooled broth or cooled around 55-65C (not solidified) agar. Swirl to mix.


The ampicillin in the diluted mixture will degrade over time. Make fresh media every 3-4 weeks


LBkan :LB-Kanamycin: add 7.5 ml of a10 mg/ml kanamycin


LBtetra: LB_tetracycline: add 1.5 ml of a 10mg/ml tetracycline



Potato Dextrose Agar (PDA)


PDA is used for the routine cultivation and identification of fungi.


Potato dextrose agar (Difco) 39 g

Distilled water 1000 ml




1. Soak potato dextrose agar in 950 of the water in a 2 L. flask. Bring to the boil, stirring constantly.


2. Autoclave at 121C for 15 minutes. Remove and pour on plates as required (sick plates fungi eat a lot!)



PDA Hygromycin 50


make 1 lt PDA , autoclave, cool down to 65C and add 130 ul of HYgromycin B (Fridge 1).

LB Broth (LB; Luria-Bertani)


10 g tryptone (bacto)

5 g yeast extract (bacto)

5 g NaCl

700 ml of nanopure water

Add contents to flask.Add stir bar and stir till dissolved

Adjust PH to 7 with NaOH. Pour into graduated cylender, Bring up to 1L volume with nanopure H2O Autoclave.


Water Agar

2% Water Agar for Plates

(from Farman Lab)



1. Agar 2% (2g of bacto Agar/100mL)

2. Pure Water (As much as needed)


3. 1L Flask

4. Foil/Autoclave tape

5. Sterile petrie plates




1. Mix water and agar in flask.

2. Autoclave on cycle 1

3. when cool enough, pour into dishes (in the fume hood). Plate thing dishes.

4. Allow to solidify

5. Allow moisture to escape by removing lids for 10-30 min.

5. Replace lids and return plates to original bags and place in the refridgerator


Regeneration medium 200 mL

(From the protocol of Bob Hanau)


amount final conc. ingredient


68.4 g 1M sucrose


3 g 1.5% agar


0.2 g 0.1% Yeast Extract


0.1 g 0.05% casein hydrolysate (enzymatic)


0.1 g 0.05% casein hydrolysate (acid)


160 mL H2O


To avoid boilovers in the autoclave, prepare in a 1 L Erlenmyer flask and completely dissolve the sucrose before autoclaving. Add Hygromycin just before using the medium


128 mL 250 mg/mL Hygromycin B (390mg/mL stock solution


Fries’ Medium


Fries’ Medium (Complete Media: CM)


Sucrose, 30g


Ammonium tartrate, 5g


Ammonium nitrate, 1g


Potassium phosphate (monobasic), 1g


Magnesium sulfate (7 H2O), 0.48g


Sodium chloride, 1g


Calcium chloride (2 H2O), 0.13g


Yeast extract, 1g


Make up to 1 liter using ultrapure water (divide in 2 bottles of 1 liter). Autoclave


if Media is for plates: Add 15 gr of bacto agar


Minimal Media (MM)

Fries’ Medium (without the Yeast extract).

if Media is for plates: Add 15 gr of bacto agar

SOC medium

To 950 mls of ultrapure water, add


20g bacto-tryptone

5g bacto-yeast extract

0.5g NaCl


Mix until dissolved. Also, make up a 1M solution of glucose (18 grams of glucose in 90 mls of ultrapure water). Autoclave both solutions separately. Allow the medium to cool to 60 degrees or less, then add 20 ml of the sterile glucose solution.


This entry was posted in Protocols. Bookmark the permalink.