User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/18

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(Procedure)
(Procedure)
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* Next, two of the erlenmeyer flasks and two of the Fernbach flasks were placed in the [[AU_Biomaterials_Design_Lab:Protocols/Autoclave|Autoclave]] at a temperature of 121°C for 60minutes followed by a 30 minute cool down period. After which, the other four samples were placed in the autoclave for 50 minutes at the same temperature and were cooled down for 30 minutes as well. The autoclave is used in order to kill all other living organisms so that they do not react with the LB because we want to only grow/culture the E.Coli cells.  
* Next, two of the erlenmeyer flasks and two of the Fernbach flasks were placed in the [[AU_Biomaterials_Design_Lab:Protocols/Autoclave|Autoclave]] at a temperature of 121°C for 60minutes followed by a 30 minute cool down period. After which, the other four samples were placed in the autoclave for 50 minutes at the same temperature and were cooled down for 30 minutes as well. The autoclave is used in order to kill all other living organisms so that they do not react with the LB because we want to only grow/culture the E.Coli cells.  
-
* The antibiotic Kanamycin was added into each of the flasks.
+
* The antibiotic Kanamycin was added into each of the flasks. The stock solution of the antibiotic was 50mg/mL. The desired concentration of kanamycin for the flasks was 50μg/mL.
-
 
+
Using the formula M<sub>1</sub>V<sub>1</sub>=M<sub>2</sub>V<sub>2</sub>
 +
The volume or amount of kanamycin that would need to go into each of the flasks was 50 μg/mL x 35mL ÷50,000 μg
 +
= .35mL Kanamycin.
 +
   
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Revision as of 04:34, 26 September 2012

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Day 5

  • Protein Expressions and Starters

Goals

  • Grow E.Coli cells and have them express ADA protein.
  • Follow Protocols for making starter and expression cultures of the cells.

Procedure

(note that while there were the amounts measured out- the amount of LB added to the flasks was actually less due to loss of sample in transferring it from the weigh dish to the flask)


(note that while there were the amounts measured out- the amount of LB added to the flasks was actually less due to loss of sample in transferring it from the weigh dish to the flask)


  • Next, two of the erlenmeyer flasks and two of the Fernbach flasks were placed in the Autoclave at a temperature of 121°C for 60minutes followed by a 30 minute cool down period. After which, the other four samples were placed in the autoclave for 50 minutes at the same temperature and were cooled down for 30 minutes as well. The autoclave is used in order to kill all other living organisms so that they do not react with the LB because we want to only grow/culture the E.Coli cells.
  • The antibiotic Kanamycin was added into each of the flasks. The stock solution of the antibiotic was 50mg/mL. The desired concentration of kanamycin for the flasks was 50μg/mL.

Using the formula M1V1=M2V2 The volume or amount of kanamycin that would need to go into each of the flasks was 50 μg/mL x 35mL ÷50,000 μg = .35mL Kanamycin.



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