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

Project name

Main project page Previous entry      Next entry

Transformation of light sensor parts The light sensor part consistutes of: BBa_B0034, Ampicillin resistence BBa_B0015, Ampicillin/Kanamycin resistence BBa_I15010, Kanamycin resistence BBa_I15009,Kanamycin resistence BBa_I15008,Kanamycin resistence BBa_R0040,Ampicillin resistence BBa_I13453,Ampicillin resistence The plasmids are transformed with competent cell, strain XL-1-Blue. Fill up the box with ice. Take 7 eppendorf tubes containing competent cells, XL-1-Blue from the -80°C freezer and straight into 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 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. Take 7 eppendorf tubes into 42°C waterbath and leave it for 1 min exactly Add 250μL of Soc into each 7 eppendorf tubes. The tubes are then left in the shaker for 1 hr Take 7 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 7 plates. Add glass beads. Now take all 7 plates and shake for 8-20 sec Remove the glass beads 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: Add 5g of bactotypton Add 0.5g of sodium chloride Add 1g of bacto-yeast extract Add 0.0327g of potassium chloride Add 200 ml of distilled water Autoclave, standard media To make Magnesium chloride: Add 0.2033 g of Mgcl2 Add 100 ml of distilled water Autoclave To make D-glucose: Add 0.182 g, D-glucose Add 100 ml of distilled water Autoclave, sugar run