User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/06: Difference between revisions
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==Purpose== | ==Purpose== | ||
* Protein was extracted from transfected BL21(DE3) E.coli cells and adenosine deaminase was purified via column chromatography. | |||
* Adenosine deaminase plasmid with the mutation E34K was transformed into BL21(DE3) E.coli cells with a second attempt. | |||
* UV-vis spectroscopy was used to analyze Au/Lysozyme solutions made on [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/10/31|2012/10/31]] to measure absorbance of supernatant of Au/Lysozyme samples | |||
==Procedure for Protein Extraction and Purification from BL21(DE3) Cells== | |||
* Frozen BL21(DE3) E.coli cells from [[User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/04|2012/11/04]] were taken out from the freezer with gloves. | |||
* The frozen cells were placed into a 500mL beaker filled with ~30°C of warm tap water for 10 minutes. After 10 minutes, cells are completely thawed and appeared in liquid form. | |||
* On ice bath, the cells were sonicated in three 1 minute intervals with three repeated cycles of procedures below: | |||
** Cells are sonicated under 8mV for 30 seconds with ______Sonicator. | |||
** Cells are immediately placed on ice for 30 seconds | |||
* After sanitation, the E.coli cells were transferred from a 25mL falcon tube into a 30mL centrifuge tube. Another 30mL centrifuge tube was filled with water with weight equal to the weight of 30mL centrifuge tube containing cellular sample. | |||
* | * | ||
==Result for Protein Extraction and Purification from BL21(DE3) Cells== | |||
{| {{table}} | {| {{table}} |
Revision as of 23:26, 23 November 2012
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Purpose
Procedure for Protein Extraction and Purification from BL21(DE3) Cells
Result for Protein Extraction and Purification from BL21(DE3) Cells
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