User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/11/06: Difference between revisions

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==Procedure==
==Procedure==
* Please refer to the [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/26| protocol]] that was previously employed. No changes were made in this procedure. 3 fracs of ADA were collected. Frac 2 had the most protein present with small amounts present in fracs 1 and 3.  
* Please refer to the FPLC [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/09/26| instructions]] that was previously employed. No changes were made in this procedure. 3 fracs of ADA were collected. Frac 2 had the most protein present with small amounts present in fracs 1 and 3. The only difference this time was that because there were not two separate samples of protein, everything could be run through the FPLC at once. the flow rate was 5mL/minute
* The transformation and plating of PCR product that was done on [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/10/24| 2012/10/24]] was unsuccessful and had to be repeated. the Original protocol for PCR mutation can be found [[AU Biomaterials Design Lab:Protocols/PCR Mutation Protocol| here]]. The following modifications were made:  
* The transformation and plating of PCR product that was done on [[User:Puja Mody/Notebook/Chem 571: Gold Nanoparticles/2012/10/24| 2012/10/24]] was unsuccessful and had to be repeated. the Original protocol for PCR mutation can be found [[AU Biomaterials Design Lab:Protocols/PCR Mutation Protocol| here]]. The following modifications were made:  
**20μL DNA solution was added to 30μL solution with E. Coli cells.
**20μL DNA solution was added to 30μL solution with E. Coli cells.

Revision as of 12:34, 24 November 2012

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FPLC and Transformation

Goals

  • Run Fast Protein Liquid Chromatography (FPLC)
  • Redo cell transformation

Procedure

  • Please refer to the FPLC instructions that was previously employed. No changes were made in this procedure. 3 fracs of ADA were collected. Frac 2 had the most protein present with small amounts present in fracs 1 and 3. The only difference this time was that because there were not two separate samples of protein, everything could be run through the FPLC at once. the flow rate was 5mL/minute
  • The transformation and plating of PCR product that was done on 2012/10/24 was unsuccessful and had to be repeated. the Original protocol for PCR mutation can be found here. The following modifications were made:
    • 20μL DNA solution was added to 30μL solution with E. Coli cells.
    • 250μL Lysogeny Broth (LB) was used instead of SOC medium.
    • Single 200μL plates were prepared. No 50μL plates were made.

Conclusions


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