User:Hana Benzeer/Notebook/SGM Summer Project/2011/06/27

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


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Transformation of light sensor parts
The light sensor part consistutes of: The plasmids are transformed with competent cell, strain XL-1-Blue.
 * 1) BBa_B0034, Ampicillin resistence
 * 2) BBa_B0015, Ampicillin/Kanamycin resistence
 * 3) BBa_I15010, Kanamycin resistence
 * 4) BBa_I15009,Kanamycin resistence
 * 5) BBa_I15008,Kanamycin resistence
 * 6) BBa_R0040,Ampicillin resistence
 * 7) BBa_I13453,Ampicillin resistence
 * 1) Fill up the box with ice.
 * 2) Take 7 eppendorf tubes containing competent cells, XL-1-Blue from the -80°C freezer and straight into 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 from each plasmid and transfer into each 7 competent cells. 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 7 eppendorf tubes into 42°C waterbath and leave it for 1 min exactly
 * 7) Add 250μL of Soc into each 7 eppendorf tubes.
 * 8) The tubes are then left in the shaker for 1 hr
 * 9) Take 7 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 7 plates.
 * 13) Add glass beads.
 * 14) Now take all 7 plates and shake for 8-20 sec
 * 15) Remove the glass beads
 * 16) The plates are then left overnight in incubater at 37°C

Results: show pic

Making Soc
Three flasks are made, flask 1 Sob, flask 2 Magnesium chloride, flask 3 D-glucose To make Sob: To make Magnesium chloride: To make D-glucose:
 * 1) Add 5g of bactotypton
 * 2) Add 0.5g of sodium chloride
 * 3) Add 1g of bacto-yeast extract
 * 4) Add 0.0327g of potassium chloride
 * 5) Add 200 ml of distilled water
 * 6) Autoclave, standard media
 * 1) Add 0.2033 g of Mgcl2
 * 2) Add 100 ml of distilled water
 * 3) Autoclave
 * 1) Add 0.182 g, D-glucose
 * 2) Add 100 ml of distilled water
 * 3) Autoclave, sugar run


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