G418 medium preparation: Difference between revisions

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<h3>Materials:</h3>
== Materials ==
<li>2x 15 ml FALCON tube
* 2x 15 ml FALCON tube
<li>filter 0.22 µl
* filter 0.22 µl
<li>5 ml syringe
* 5 ml syringe
<li>DMEM
* DMEM
<li>G418
* G418


<h3>Execution:</h3>
== Procedure ==
# 1 g of weight has a total concentration from 0,751 g
# 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?
# How much of DMEM we need for 1,5 g G418 if we prepare a stock solution with a concentration of 200 mg/ ml?

Revision as of 09:16, 25 March 2015

Materials

  • 2x 15 ml FALCON tube
  • filter 0.22 µl
  • 5 ml syringe
  • DMEM
  • G418

Procedure

  1. 1 g of weight has a total concentration from 0,751 g
  2. How much of DMEM we need for 1,5 g G418 if we prepare a stock solution with a concentration of 200 mg/ ml?
  3. 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