Difference between revisions of "Team:UCLA/Notebook/Recombinant Expression/3 July 2015"
| Line 292: | Line 292: | ||
Preparation of 1X Lysis Buffer: | Preparation of 1X Lysis Buffer: | ||
5 mL of 8X Lysis Buffer | 5 mL of 8X Lysis Buffer | ||
| − | 35 mL of DI | + | 35 mL of DI <sup>superscript</sup> |
Revision as of 22:53, 9 July 2015
Beginning of Cell Lysis
The start-up culture incubated overnight in 1 L of LB is removed from the shaking incubator. An OD check is performed, and reported to be 0.65. The contents from the flask are divided up into 20 Falcon tubes, with each one containing approximately 45 mL of start-up culture in LB. The tubes are centrifuged for 15 minutes at 5,000 g at 4 degrees Celsius. The liquid is decanted and the cell pellet is weighed. The cell pellet is then suspended in lysis buffer in a 1 to 3 weight (g) to volume (mL) ratio. The tubes are stored at -80 degrees Celsius.
Preparation of 1X Lysis Buffer: 5 mL of 8X Lysis Buffer 35 mL of DI superscript