Media protocol for DH5 Alpha?

Question

Hi

Does anyone has a protocol of the preparation of medium for DH5 Alpha bacteria cells for OPTIMUM growth?

Answer 1

It depends what you need the bacteria for.

For normal expansion, they grow very well in normal LB medium

However, if you want to grow your bacteria in order to make competent cells, then you should use SOB or 2YT media and transfrom them using SOC.

Here are the recipies for these 3 media:

Luria Bertani Medium (LB)
(Makes 1000 mL)

10 g/L Bacto -Tryptone, 5 g/L Bacto -Yeast Extract, 5 g/L NaCl, adjust the pH to 7.5 with NaOH autoclave to sterilize. Allow the auto-claved medium to cool to 55ºC and add ampicillin (final concentration 100 µg/ml). For LB plates, 1.5% Bacto-agar (15 g/L) was added prior to autoclaving.

S.O.C. medium.

2% Tryptone (bacto), 0.5% Yeast Extract, 10 mM NaCl, 2.5 mM MgCl2, 10 mM MgSO4, 20 mM glucose. Dissolve the reagents by stirring on a magnetic stirrer. Adjust the pH to 7.0 with approximately 0.2 ml of 5 N NaOH. Sterilize by autoclaving.

S.O.B. medium
(Makes 1000ml)

20 g Peptone from casein , 5 g Yeast Extract, 0.5 g NaCl (cat.no. 30183)
Dissolve the reagents by stirring on a magnetic stirrer. Adjust the pH to 7.0 with approximately 0.2 ml of 5 N NaOH.
Sterilize by autoclaving. Just before use add: 2.5 mM KCl , 10 mM MgCl2 , 10 mM MgSO4

2 x YT Medium
(Makes 1000ml)
6 g Peptone from casein, 10 g Yeast Extract , 5 g NaCl
Dissolve the reagents by stirring on a magnetic stirrer. Adjust the pH to 7.0 with NaOH. Fill up to a final volume of 1 liter with deionized water. Sterilize by autoclaving.

Answer 2

Dh5 Alpha

Answer 3

Dh5alpha