Difference between revisions of "G418 medium preparation"

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(Created page with "'''Materials:''' - 2x 15 ml FALCON tube - filter 0.22 µl - 5 ml syringe - DMEM - G418 '''Execution:''' 1 g of weight has a total concentration from 0,751 g How much of DMEM ...")
 
 
(8 intermediate revisions by the same user not shown)
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'''Materials:'''
+
== Material ==
- 2x 15 ml FALCON tube
+
* 2x 15 ml FALCON tube
- filter 0.22 µl
+
* filter 0.22 µl
- 5 ml syringe
+
* 5 ml syringe
- DMEM
+
* DMEM
- G418
+
* G418
  
'''Execution:'''
+
== 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?
1.5g powder has a total concentration from 1,1256 g G418
+
# 1.5g powder has a total concentration from 1,1256 g G418
  
 
1125,6 mg/ 200 mg = 5,6  
 
1125,6 mg/ 200 mg = 5,6  
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
+
* dissolve the powder in DMEM
- if it is dissolve than filter the solution:
+
* 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
+
* 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
  
for using:
+
<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
+
* 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. 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