User:Hana Benzeer/Notebook/SGM Summer Project/2011/06/07
Transformation of four strains of competent cell: DH5α, JMV 101, MACH1 and XL-1BLUE with plasmid BBa_Kate_GFP
- Take out DH5α strainfrom the -80°C freezer and place it in the ice bucket. Repeat this for other strains.
- Take out plasmid BBa_Kate_GFP from the -20°C freezer and plave it in the ice bucket.
- Take out the Soc from the -20°C freezer and place it in the incubater at 37°C.
- Add 1μL of the plasmid and transfer into each strains. Mix it very well. Note: This need to be completed in front of the flame.
- Put the tubes back into the ice bucket for 5 min.
- The tubes is placed in waterbath at 42°C for 1 min exactly.
- Add 250μL of Soc into each tubes and vortex.
- The tubes is then placed in the shaker for about 1 hr.
- Take out agar plates with Ampicillin and place it in the incubater at 37°C for 5 min upsidedown with the lid open slightly.
- Take out the tubes from the shaker and the plates from the incubater.
- Transfer the strains into each plates by pouring onto the surface of the plates.
- Add 8-10 glass beads into the plate and shake the plates side to side and all around.
- Remove the glass beads by tapping on one side of the plate.
- The plates are placed in the incubater at 32°C for overnight (16hrs).
Four 10 ml bottles of LB Broth with Ampicillin were inoculated and grown overnight from colonies generated from various strains transformed with plasmid BBa_Kate_GFP
- Four 10 ml LB broth bottles are required for four 200 ml plates that are transformed overnight with plasmid BBa_Kate_GFP.
- The colonies are removed from each plates using----- and inoculated into each 10 ml broth tube.
- The bottles are labelled and placed in the shaker for 16 hrs, overnight.