User:Michael F. Nagle/Notebook/Chem 571/2012/11/06

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Current revision (06:33, 7 December 2012) (view source)
(Procedure)
 
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==Procedure==
==Procedure==
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*The prodedure for [AU_Biomaterials_Design_Lab:Protocols/PCR_Mutation_Protocol|PCR Mutation] was followed, with the following modifications
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*The prodedure for [[AU_Biomaterials_Design_Lab:Protocols/Transformation_Protocol|transformation]] was followed, just like on [[User:Michael_F._Nagle/Notebook/Chem_571/2012/10/24|10/24]] but with the following modifications
**20μL DNA solution was added to 30μL solution with E. Coli cells.
**20μL DNA solution was added to 30μL solution with E. Coli cells.
**250μL Lysogeny Broth (LB) was used instead of SOC medium.
**250μL Lysogeny Broth (LB) was used instead of SOC medium.
**Single 200μL plates were prepared. No 50μL plates were made.
**Single 200μL plates were prepared. No 50μL plates were made.
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*Adenosine Deaminase prepared [User:Michael_F._Nagle/Notebook/Chem_571/2012/10/23|Oct. 23] was purified via FPLC
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**Plates were incubated at 37<sup>o</sup>C
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*Adenosine Deaminase prepared [[User:Michael_F._Nagle/Notebook/Chem_571/2012/10/23|Oct. 23]] was purified via FPLC [[User:Michael_F._Nagle/Notebook/Chem_571/2012/09/26|as was done 9/26]]
** Pump flow was 5mL/minute
** Pump flow was 5mL/minute

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Objectives

  • Transform Adenosine Deaminase in cells
  • Purify protein via Fast Protein Liquid Chromatography (FPLC)

Procedure

  • The prodedure for transformation was followed, just like on 10/24 but with the following modifications
    • 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.
    • Plates were incubated at 37oC
  • Adenosine Deaminase prepared Oct. 23 was purified via FPLC as was done 9/26
    • Pump flow was 5mL/minute



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