G418 medium preparation: Difference between revisions
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== | == Material == | ||
* 2x 15 ml FALCON tube | * 2x 15 ml FALCON tube | ||
* filter 0.22 µl | * filter 0.22 µl | ||
| Line 15: | Line 15: | ||
it means, that you need 5,6 ml DMEM for 1,5 g G418 , if you wanted a concentration of 200 mg/ ml | it means, that you need 5,6 ml DMEM for 1,5 g G418 , if you wanted a concentration of 200 mg/ ml | ||
* dissolve the powder in DMEM | |||
* if it is dissolve than filter the solution: | |||
put the pump from the syringe out, fill the antibiotic in the syringe, take the pump back and press all across the filter | put the pump from the syringe out, fill the antibiotic in the syringe, take the pump back and press all across the filter | ||
* label the vessel with the name of the antibiotics and the stock solution | |||
f.e. G418 200 mg/ ml in DMEM | f.e. G418 200 mg/ ml in DMEM | ||
<h5>for using:</h5> | <h5>for using:</h5> | ||
HS with antibiotics: 400 µg/ ml | HS with antibiotics: 400 µg/ ml | ||
* 0,2/ 0,0004 = 500 ⇒ 100 ml media/ 500 ⇒ 0,2 ml antibiotics in 1200 ml media | |||
Latest revision as of 14:19, 8 June 2015
Material
- 2x 15 ml FALCON tube
- filter 0.22 µl
- 5 ml syringe
- DMEM
- G418
Procedure
- 1 g of weight has a total concentration from 0,751 g
- How much of DMEM we need for 1,5 g G418 if we prepare a stock solution with a concentration of 200 mg/ ml?
- 1.5g powder has a total concentration from 1,1256 g G418
1125,6 mg/ 200 mg = 5,6
it means, that you need 5,6 ml DMEM for 1,5 g G418 , if you wanted a concentration of 200 mg/ ml
- dissolve the powder in DMEM
- if it is dissolve than filter the solution:
put the pump from the syringe out, fill the antibiotic in the syringe, take the pump back and press all across the filter
- label the vessel with the name of the antibiotics and the stock solution
f.e. G418 200 mg/ ml in DMEM
for using:
HS with antibiotics: 400 µg/ ml
- 0,2/ 0,0004 = 500 ⇒ 100 ml media/ 500 ⇒ 0,2 ml antibiotics in 1200 ml media