User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/10/24

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(Procedure)
(Procedure)
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==Procedure==
==Procedure==
* The PCR product was transformed by following the [[AU Biomaterials Design Lab:Protocols/Transformation Protocol| Transformation Protocol]]
* The PCR product was transformed by following the [[AU Biomaterials Design Lab:Protocols/Transformation Protocol| Transformation Protocol]]
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The only changes that were made are as follows:
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# Of the two cell solutions made, 200μL of  LB media was added to one of the solutions while 200 μL of SOC media was added to the other solution of cells. The two solutions were then shaken at 275 rpm for 1 hr.
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#25 μL of transformed cells was plated on one LB Petri dish, while 200 μL of transformed cells was plated on the other. For K110A mutation, a total of 4 plates were made. 2 for K110A(f) and 1 for K110A (r).
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* ADA protein was expressed in the same manner as on [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/19| 2012/09/19]]
* ADA protein was expressed in the same manner as on [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/19| 2012/09/19]]
** 5mL of .4M IPTG solution was made and 1mL was placed into each of the 4 fernbach flasks.  
** 5mL of .4M IPTG solution was made and 1mL was placed into each of the 4 fernbach flasks.  

Revision as of 19:08, 21 November 2012

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Transformation, protein expression, lysozyme Uv-Vis

Goals

  • Transform PCR product into E.Coli Cells
  • Induce ADA protein expression
  • Run Uv-Vis of the lysozyme and AuNP solution

Procedure


The only changes that were made are as follows:

  1. Of the two cell solutions made, 200μL of LB media was added to one of the solutions while 200 μL of SOC media was added to the other solution of cells. The two solutions were then shaken at 275 rpm for 1 hr.
  2. 25 μL of transformed cells was plated on one LB Petri dish, while 200 μL of transformed cells was plated on the other. For K110A mutation, a total of 4 plates were made. 2 for K110A(f) and 1 for K110A (r).
  • ADA protein was expressed in the same manner as on 2012/09/19
    • 5mL of .4M IPTG solution was made and 1mL was placed into each of the 4 fernbach flasks.
    • .005L x.4M= .002 moles x 238.31g/mol= .477g IPTG in 5mL. Actual= .4754g in 5mL of water.
  • For information regarding the Lysozyme please see either Mikes notebook or Marys notebook. The lysozyme did create purple gold nanoparticle fibers upon removal from the oven.




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