User:Hana Benzeer/Notebook/SGM Summer Project/2011/06/28

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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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Inoculation
Only 5 parts were used to incoulate, BBa_I13453, BBa_B0034, BBa_I15009, BBa_R0040 and BBa_B0015
 * 1) 2 10ml tubes for each (10 total)
 * 2) Label the tubes
 * 3) Turn on the flame
 * 4) I13453, add 10μL Ampicillin
 * 5) B0034, add 10μL Ampicillin
 * 6) I15009, add 50μL Kanamycin
 * 7) R0040, add 10μL Ampicillin
 * 8) B0015, add 10μL of Ampicillin and 50μL of Kanamycin
 * 9) Inoculate the 5 plates into each 2x 10 ml tubes.
 * 10) 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.
 * 1) Fill up the box with ice.
 * 2) Take 4 eppendorf tubes, 2x XL-1-Blue and 2x DH5-α from the -80°C freezer and put it on ice.
 * 3) 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
 * 4) Take out the Soc from the -20°C freezer°C and leave in the incubater for 10 min
 * 5) 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.
 * 6) Take 4 eppendorf tubes into 42°C waterbath and leave it for 1 min exactly
 * 7) Add 250μL of Soc.
 * 8) The tubes are then left in the shaker for 1 hr
 * 9) Take 4 selective agar plates and leave it in the incubater upside down with lid open slightly to lit air through.
 * 10) Take out the tubes from the shaker
 * 11) Take out the plates from the incubater.
 * 12) Turn on the flame. Transfer the contents from each epppendorf tubes into each 4 plates.
 * 13) Add glass beads.
 * 14) Now take all 4 plates and shake for 8-20 sec
 * 15) Remove the glass beads
 * 16) The plates are then left overnight in incubater at 37°C


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