User:Keyun Wang/Notebook/Experimental Biological Chemistry I/2012/11/04: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Experimental Biological Chemistry I</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Experimental Biological Chemistry I</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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==Purpose== | ==Purpose== | ||
* Grow and amplify ADA transfected BL21(DE3) E.coli cells in LB media | * Grow and amplify ADA transfected BL21(DE3) E.coli cells in LB media | ||
* Extract cells from media after ADA protein expression | * Extract cells from media after ADA protein expression | ||
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* The absorbance for cells after 4 hours of growth were higher than expected (an absorbance of 0.600 at 600nm). This indicates the cells were overgrown. The cells were harvested nonetheless. The high absorbance indicates more cells in flasks, thus more protein when purified. | * The absorbance for cells after 4 hours of growth were higher than expected (an absorbance of 0.600 at 600nm). This indicates the cells were overgrown. The cells were harvested nonetheless. The high absorbance indicates more cells in flasks, thus more protein when purified. | ||
* The cell cultures were centrifuged for 30 minutes as oppose to the protocol which suggests 15 minutes. Longer centrifugation time is used here to make sure all cells were in the pellet. | * The cell cultures were centrifuged for 30 minutes as oppose to the protocol which suggests 15 minutes. Longer centrifugation time is used here to make sure all cells were in the pellet. | ||
* The final volume of cell resuspension is 42mL due to the addition of cell pellet as well as small amounts of supernatant that were not completely cleared out. | |||
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__NOTOC__ | __NOTOC__ |
Latest revision as of 22:11, 26 September 2017
Experimental Biological Chemistry I | Main project page Previous entry Next entry | ||||||||||||||||||||
Purpose
Procedure
Molecular weight: 238.31g/mol 0.4M × 0.004L = 0.0016mol of IPTG needed 238.31g/mol × 0.0016mol = 0.3813g of IPTG needed
Wavelength: 600nm Settings: Photometric Data display: Absorbance
20mM Tris buffer 0.5M NaCl 30mM Imidizole pH 7.5
Notes
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