DH5α Incubation in Liquid Media 6/28/2013
- With a pipetter, 3mL of LB+amp was added to a test tube.
- A colony was selected on the culture plate(plate was overgrown), and a circle was drawn where the colony was selected.
- Micropipette tip was lightly touched to the colony making sure not to scrape any of the agar gel. Micropipette tip was then ejected into test tube.
- Test tube was incubated with shaker for 7 hours at 37 degrees.
DH5α Mini Prep with KAH013 6/28/2013
- Colonies that Rene had incubated in liquid media on 6/27 were used.
- Using a micropipette, a 2mL centrifuge tube was filled with the incubated liquid media.
- The solution was centrifuged at 13.1g for 3 minutes (If a pellet has not completely condensed centrifuge again until pellet does condense).
- Using a micropipette, the supernatant was removed until 600μL of solution remained in the centrifuge tube. (Discard supernatant into the bleach container in the fume hood)
- Pellet was broken up using the vortex.
- Added 100μL of lysis buffer to the centrifuge tube and inverted 4-6 times and waited 1 min.
- Added 350μL of lysis buffer. Invert the solution until it turns yellow, make sure no blue pockets are left.
- Centrifuge the solution at