Preparation procedure of LB medium

February 13 05:06:23, 2023

Experimental procedure

LB medium
Tryptone
10 g
Yeast Extract
5 g
NaCl
10 g
Add water to 1000 mL (pH 7.2, solid medium plus 1.8% agar powder).
2. LB resistance screening medium
The temperature of the LB solid medium to be sterilized is reduced to about 50 °C, and the antibiotic stock solution is added to a final concentration of 50 μg/mL, that is, 1 μL of the antibiotic stock solution is added per ml of the medium, shaken, and then the plate is inverted.
The liquid LB resistant screening medium was added with antibiotics after complete cooling, in the same amount as the solid medium.
3. LB/Amp/IPTG/X-gal medium
Add 40 μL of X-gal stock solution and 40 μL of IPTG solution to LB agar plates containing 100 μg/mL Amp, apply the solution evenly to the entire surface of the plate with a sterile glass applicator, and dry on a clean bench. After the backup.
4. SOC liquid medium
Tryptone
20 g
Yeast Extract
5 g
NaCl
0.5 g
Add 950 mL of water, completely dissolve, add 10 mL of 250 mmol/L KCl solution, adjust the pH to 7.0 with NaOH solution (5 mmol/L), dilute to 1000 mL with double distilled water, and autoclave. After sterilization, cool down to below 60 ° C and add 7.2 mL of 50% glucose solution. Add 5 mL of sterilized 2 mol/L MgCl 2 solution before use.

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