User:Dhea Patel/Notebook/Experimental Biological Chemistry Notebook/2012/11/06

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Current revision (16:45, 6 December 2012) (view source)
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Objective

  • to run transformations of PCR product into E. coli cells
  • to run FPLC on the ADA protein from the third and the fourth of November.

Description

  • The cells from the fourth of November were sonciated for 30 seconds and then placed in an ice bath for 30 seconds. This was repeated for a total of 3 times.
  • The cells were then transferred to centrifuge tubes, balanced, and placed in the centrifuge at 18,000rpm at 4°C for 2 hours.
  • The cells were then filtered using Supor®-450 47mm membrane filter.
  • The cells were then run on FPLC, using the binding buffer and elusion buffer from September 26th.
  • The cells were collected in 5 mL aliquots and transferred into a Falcon tube.


  • The Au/Lysozyme solutions were obtained from the oven. The following images are the results from the Au/Lysozyme reaction.

Image:IMAG0851.jpg

  • The range of Au/Lysozyme ratios, between 20uM ratio to 130 uM ratio.

  • A close up of Au/Lysozyme ratios, focusing on the difference between the homologous solutions and the protein fiber solutions.
  • UV-Vis was run on the Au/Lyzozyme solutions.
  • For the transforamtion of PCR product into E. coli cells, 20μL of PCR DNA and 30μL of E. coli cells were added to a centrifuge tube, in a sterile environment, and then placed in an ice bucket. (This occurred twice, one for each PCR tube.)
  • The cells were heat schoked by incubating them in a 42°C water bath for 30 seconds and then placed immediately on ice.
  • 250μL of room temperature SOC medium was added to one of the PCR tubes and 250μL of Albumin medium was added to the other PCR tube.
  • The cells were shaken at 225 rpm of an hour at 37°C.

Data

  • This graph shows the drop off of of the concentration of Au after 60 Au:Lys. This means that gold nanoparticles were no longer in solution at 70-130 Au:Lysozyme.

Image:Percent_change_in_concentration_of_au_in_aulys_soln.png

  • This graph shows the percent change in Au concentration vs. Au:Lys mole ratio. Every sample contains gold nanoparticles in their solutions.


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