Check overnight growth (7x200ml)
Only 5 parts were used to incoulate, BBa_I13453, BBa_B0034, BBa_I15009, BBa_R0040 and BBa_B0015
- 2 10ml tubes for each (10 total)
- Label the tubes
- Turn on the flame
- I13453, add 10μL Ampicillin
- B0034, add 10μL Ampicillin
- I15009, add 50μL Kanamycin
- R0040, add 10μL Ampicillin
- B0015, add 10μL of Ampicillin and 50μL of Kanamycin
- Inoculate the 5 plates into each 2x 10 ml tubes.
- 10 tubes are then left in the shaker overnight
Transformation of BBa_I15008 and I15010
The plasmids are transformed with 2 different competent cells, XL-1-Blue and DH5-α.
- Fill up the box with ice.
- Take 4 eppendorf tubes, 2x XL-1-Blue and 2x DH5-α from the -80°C freezer and put it on ice.
- Take the plasmids (Igem plates 2010. If the plates are not used, meaning the well of interest is not puncrures, add 10μL of free nuclease water)from the -20°C freezer
- Take out the Soc from the -20°C freezer°C and leave in the incubater for 10 min
- Add 2μL, plasmid and transfer into each 4 eppendorf. Mix the DNA to the cells by vortex and leave in the ice for 5 min. Note= while doing this, flame should be on.
- Take 4 eppendorf tubes into 42°C waterbath and leave it for 1 min exactly
- Add 250μL of Soc.
- The tubes are then left in the shaker for 1 hr
- Take 4 selective agar plates and leave it in the incubater upside down with lid open slightly to lit air through.
- Take out the tubes from the shaker
- Take out the plates from the incubater.
- Turn on the flame. Transfer the contents from each epppendorf tubes into each 4 plates.
- Add glass beads.
- Now take all 4 plates and shake for 8-20 sec
- Remove the glass beads
- The plates are then left overnight in incubater at 37°C