*to run FPLC on the ADA protein from [[User:Dhea_Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/11/03| the third]] and [[User:Dhea_Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/11/04 | the fourth]].

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*to run transformations of PCR product into E. coli cells

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*to run FPLC on the ADA protein from [[User:Dhea_Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/11/03| the third]] and [[User:Dhea_Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/11/04 | the fourth]] of November.

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==Description==

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*The cells from [[User:Dhea_Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/11/04 | 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.

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*The cells were then transferred to centrifuge tubes, balanced, and placed in the centrifuge at 18,000rpm at 4°C for 2 hours.

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*The cells were then filtered using Supor®-450 47mm membrane filter.

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*The cells were then run on FPLC, using the binding buffer and elusion buffer from [[User:Dhea_Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/09/26| September 26th]].

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*The cells were collected in 5 mL aliquots and transferred into a Falcon tube.

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*The [[User:Dhea_Patel/Notebook/Experimental_Biological_Chemistry_Notebook/2012/10/31|Au/Lysozyme solutions]] were obtained from the oven. The following images are the results from the Au/Lysozyme reaction.

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[[Image:IMAG0851.jpg]]

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

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[[Image:IMAG0852.jpg|650px]]

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*A close up of Au/Lysozyme ratios, focusing on the difference between the homologous solutions and the protein fiber solutions.

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*UV-Vis was run on the Au/Lyzozyme solutions.

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*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.)

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*The cells were heat schoked by incubating them in a 42°C water bath for 30 seconds and then placed immediately on ice.

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*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.

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*The cells were shaken at 225 rpm of an hour at 37°C.

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==Data==

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[[Image:Final_concentration_of_Au_in_AuLys_sol.png|550px]]

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*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.

Current revision

Objective

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.

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.