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Transformation_Vector_info.pdf

1. Allow tube containing competent E. coli cells to thaw on ice.

2. When cells have thawed pipette 2 μl of the of plasmids into the cells.

3. Mix the DNA and cells by gently flicking the tube.

4. Incubate on ice for 5 minutes.

5. Heat-shock the cells for 30 seconds at 42°C.

6. Immediately transfer the tubes to ice.

5. Add 250 μl of room temperature LB Medium.

6. Cap the tube tightly and shake the tube at 37°C for 1 hour.

7. Spread cells onto plates as indicated in the table below.

.....Quantity of cells (µL).....................Plate type
.....2 µL.............................................LB (one line on plate)
.....2 µL.............................................LB Ampicillin (two lines on plate)
.....2 µL.............................................LB Amp, Arabinose, IPTG (three lines on plate)
.....100 µL.........................................LB Amp, Arabinose, IPTG (three lines on plate)

8. Grow overnight at 37°C.