 |
|
|
|
|
|
|
|
|
|
|
|
|
| HOME | ||||||||||||
| Isolation of Plasmid DNA from Yeast | ||||||||||||
| In order to isolate the cDNA plasmid from yeast, one colony (His+, LacZ+) was inoculated into YPAD broth and grown at 30 degrees C for 2 days. The culture was peletted at full speed, lysed and extracted DETAIL HERE . The DNA was washed with ethanol after centrifugation and re-spun at 14,000 GO AWAY rpm for 10 minutes and supernatant decanted under a vacuum. The DNA was transformed into Escheria coli cells by electroporation according to the manufacturer��s instructions (Cell-Porator, Life Technologies, Rockville, MD, USA.) The E. Coli were transferred to LB Amp-50 media and allowed to grow. FIG 6 Two colonies were picked from each plate and placed on liquid LB amp media. Colonies are transferred to filter paper on SD plates and assayed for expression of the lacZ reporter gene by the detection of B-galactosidase activity with an X-gal containing solution. The streaked plates were allowed to grow overnight and then treated with liquid nitrogen before addition of substrate. |
||||||||||||
 |
||||||||||||
| rice view | ||||||||||||
 |
||||||||||||
| yeast | ||||||||||||