Transformation

From 2007.igem.org

(Difference between revisions)
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1. To 50uL competent cells add 1uL DNA solution
1. To 50uL competent cells add 1uL DNA solution
 +
2. Incubate on ice for 45min
2. Incubate on ice for 45min
 +
3. Heat shock at 42 degrees for 1min
3. Heat shock at 42 degrees for 1min
 +
4. Incubate on ice for 15min
4. Incubate on ice for 15min
 +
5. Add 1ml LB
5. Add 1ml LB
 +
6. Incubate at 37 degrees for 1 hour
6. Incubate at 37 degrees for 1 hour
 +
7. Pellet Cells
7. Pellet Cells
 +
8. Pour off majority of LB
8. Pour off majority of LB
 +
9. Resuspend cells in remaining LB
9. Resuspend cells in remaining LB
 +
10.Plate resuspended cells and incubate overnight at 37 degrees
10.Plate resuspended cells and incubate overnight at 37 degrees

Revision as of 04:16, 27 June 2007

Transformation

1. To 50uL competent cells add 1uL DNA solution

2. Incubate on ice for 45min

3. Heat shock at 42 degrees for 1min

4. Incubate on ice for 15min

5. Add 1ml LB

6. Incubate at 37 degrees for 1 hour

7. Pellet Cells

8. Pour off majority of LB

9. Resuspend cells in remaining LB

10.Plate resuspended cells and incubate overnight at 37 degrees