media/plates

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David E
David E's picture
media/plates

Ampicillin (Amp): Stock of 5 mg/ml in sterile ddwater (sddH2O).
0.5 g Amp
sddH2O to 100 ml (Add to media for final conc. 100 ug/ml)

Lambda plates:
10 g Bacto-tryptone
15 g Bacto-agar
2.5 g NaCl
ddH2O to 1 L
autoclave to sterilize and pour into sterile petri dishes (approx. 20 ml/plate).

Lambda top agar:
10 g Bacto-tryptone
10 g Bacto-agar
5 g NaCl
ddH2O to 1 L
autoclave to sterilize

LB Medium:
10 g Bacto-Tryptone
5 g Bacto-yeast extract
10 g NaCl
ddH2O to 1 L
autoclave to sterilize

LB plates:
10 g Bacto-Tryptone
5 g Bacto-yeast extract
10 g NaCl
15 g Bacto-agar
ddH2O to 1 L

autoclave to sterilize, cool to 55deg.C, add antibiotic if desired, and pour into sterile petri dishes (approx. 20 ml/plate).

M-9 Agar:
Add 15 g. agar to 870 ml. double distilled water in a 2 L Ehrlenmeyer flask and autoclave. Also autoclave a 100 ml. graduate cylinder capped with aluminum foil to use for measuring the sterile M-9 salts later. Swirl the agar gently and carefully upon removal from autoclave to disperse any undissolved agar. Allow to cool in a 55degC water bath. When 55degC, add the ingredients called for in the M-9 liquid medium recipe, omitting the water. Be sure to use sterile pipettes or graduate cylinders, as this mixture cannot be autoclaved. Immediately pour into sterile Petri dishes, using sterile technique.

M-9 Medium (liquid):
100 ml 10X M-9 salts
1 ml 1 M MgSO4 (autoclaved)
10 ml 20% glucose (filter sterilized),
1 ml 1% thiamine (filter sterilized)
10 ml 100 mM CaCl2 (autoclaved)
sddH2O to 1 L

10X M-9 Salts:
60 g Na2HPO4 (sodium phosphate, dibasic)
30 g KH2PO4 (potassium phosphate, monobasic)
5 g NaCl
10 g NH4Cl (ammonium chloride)
ddH2O to 1 liter (autoclave)

10xTB Salts:
2.31 g KH2PO4
12.54 g K2HPO4 (potassium phosphate, dibasic)
ddH2O to 100 ml autoclave)

Terrific Broth (TB):
12 g Bacto-tryptone
24 g yeast extract
4 ml glycerol
ddH2O to 900 ml
Autoclave, cool and add 100 ml of 10xTB Salts and adjust the final volume to 1 L with sddH2O.

Tetracycline stock (Tet): Stock of 10 mg/ml in 50% ethanol + sddwater.
1 g Tetracycline
50 ml 100% ethanol
sddH2O to 100 ml (store at 4deg.C in the absence of light)

Add to media for final conc. 20 ug/ml.

2xTY medium:
16 g Bacto-tryptone
10 g Bacto-yeast extract
5 g NaCl
ddH2O to 1 L (autoclave)

eloradana
eloradana's picture
Hi David,

Hi David,

I have a question, do you know how many time is possible storage 2xTY at 4ºC? 

Thank you in advance!