Difference between revisions of "Team:UCLA/Notebook/Recombinant Expression/18 July 2015"
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'''Weight of Empty 500mL container:''' 80.56g | '''Weight of Empty 500mL container:''' 80.56g | ||
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'''Weight of 500mL container with Cell Pellet:''' 84.06g | '''Weight of 500mL container with Cell Pellet:''' 84.06g | ||
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'''Weight of 2nd 500mL container with Cell Pellet:''' 84.55g | '''Weight of 2nd 500mL container with Cell Pellet:''' 84.55g | ||
'''Weight of Cell Pellet:''' 84.06g-80.56g = 3.5 g | '''Weight of Cell Pellet:''' 84.06g-80.56g = 3.5 g | ||
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'''Weight of 2nd Cell Pellet:''' 84.55g-80.56g = 3.99g, round to 4 g | '''Weight of 2nd Cell Pellet:''' 84.55g-80.56g = 3.99g, round to 4 g | ||
Revision as of 23:50, 18 July 2015
The flask is removed from the shaking incubator and the contents are divided into two 500mL containers. The containers are weighed and a few drops of water is added to the lighter one to ensure proper counterbalance in the centrifuge. The two containers are centrifuged at 5,000g at 4 degrees Celsius for 15 minutes. The supernatant is decanted and the cell pellets are weighed, and then suspended in 1X Lysis Buffer in a 1 to 3 weight of cell pellet to volume of buffer, respectively, ratio.
Weight of Empty 500mL container: 80.56g
Weight of 500mL container with Cell Pellet: 84.06g
Weight of 2nd 500mL container with Cell Pellet: 84.55g
Weight of Cell Pellet: 84.06g-80.56g = 3.5 g
Weight of 2nd Cell Pellet: 84.55g-80.56g = 3.99g, round to 4 g