User:Hana Benzeer/Notebook/SGM Summer Project/2011/06/07

Transformation protocol
Transformation of four strains of competent cell: DH5α, JMV 101, MACH1 and XL-1BLUE with plasmid BBa_Kate_GFP


 * 1) Take out DH5α strainfrom the -80°C freezer and place it in the ice bucket. Repeat this for other strains.
 * 2) Take out plasmid BBa_Kate_GFP from the -20°C freezer and plave it in the ice bucket.
 * 3) Take out the Soc from the -20°C freezer and place it in the incubater at 37°C.
 * 4) 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.
 * 5) Put the tubes back into the ice bucket for 5 min.
 * 6) The tubes is placed in waterbath at 42°C for 1 min exactly.
 * 7) Add 250μL of Soc into each tubes and vortex.
 * 8) The tubes is then placed in the shaker for about 1 hr.
 * 9) Take out agar plates with Ampicillin and place it in the incubater at 37°C for 5 min upsidedown with the lid open slightly.
 * 10) Take out the tubes from the shaker and the plates from the incubater.
 * 11) Transfer the strains into each plates by pouring onto the surface of the plates.
 * 12) Add 8-10 glass beads into the plate and shake the plates side to side and all around.
 * 13) Remove the glass beads by tapping on one side of the plate.
 * 14) The plates are placed in the incubater at 32°C for overnight (16hrs).

Inoculation
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


 * 1) Four 10 ml LB broth bottles are required for four 200 ml plates that are transformed overnight with plasmid BBa_Kate_GFP.
 * 2) The colonies are removed from each plates using- and inoculated into each 10 ml broth tube.
 * 3) The bottles are labelled and placed in the shaker for 16 hrs, overnight.