Commonly used solution preparation in cell culture

Commonly used solution preparation in cell culture (PBS, DMEM, glutamine, G418, etc.)

PBS

NaCl 8 g

KCl 0.2 g

KH2PO4 0.2 g

Na2HPO4 · 12H2O 3.634g

Add dH2O to 900ml, adjust pH to 7.4, replenish water to 1L, dispense into 5 blue capped bottles, and sterilize.

TP + EDTA mixed solution

Weigh TP 0.25g, EDTA 0.02g, dissolve in 200ml PBS, filter and sterilize.

DMEM and RPMI 1640

The former weighs 3.7g of NaHCO3 and the latter weighs 2.0g.

Adjust the pH to 7.2 ~ 7.4.

Preparation of glutamine

Weigh 2.922g of Gln, dissolve it in three distilled water, add to 100ml, then make up 200mM stock solution. Stir to dissolve thoroughly, filter and sterilize, divide into 2ml EP tubes, and store at -20 ℃. During use, 1ml Gln stock solution can be added to 100ml culture medium.

Because glutamine is very unstable in the solution, it can be decomposed by 50% for 1 week at 4 ° C. When the culture solution with glutamine is stored in the refrigerator at 4 ° C for more than 2 weeks, the original glutamine should be added again.

G418 stock solution (0.1 mg / uL)

Add 400uL of stock solution to 100ml medium, then the screening concentration is 400ug / ml.

Add 200 uL of stock solution to 100 ml of culture medium to maintain a concentration of 200 ug / ml.

LB liquid medium

Tryptone 10g

Yeast extract 5g

NaCl 10g

dH2O 1L

Adjust the pH to 7.0 with about 0.2 ml of 5M NaOH. Sterilize for 20min.

LB solid medium, plus 12 ~ 15g agar.

Addition of Ampicillin in LB

Weigh 5g ampicillin, add water to bring the volume to 50ml, fully dissolve, and the obtained stock solution concentration is 100mg / ml. Filter and sterilize, aliquot into EP tubes and store at -20 ℃.

To make its working concentration 100ug / ml, it needs to be diluted 1000 times.

That is, add 1ml to 1000ml medium. Or add 100ul to 100ml medium.

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