Joyce:Transformation

Procedure (for E. coli XL2-Blue)

1. Thaw cells
2. Pipette 50 ul of cells into an ependorff
3. With gentle stirring, pippette 5 ul of a 20 ul (about 100 ng? *****) ligation reaction into the ependorff
4. Place the ependorff on ice for at least 30 minutes
5. Heat shock the transformation mixture for 45 seconds at 42*C
6. Put the transformation mixture on ice for 2 minutes
7. Add 400 ul of sterile YT to the transformation mixture
8. Mix gently by inverting several times
9. Incubate at 37*C for at least 45 minutes
10. Centrifuge at full speed for 2 minutes
11. Discard 300 ul of the supernatant
12. Resuspend the pellet in the remaining 150 ul
13. Plate and incubate at 37*C overnight

Notes

• You can do a cheap-o transformation to quickly check something by incubating 100 ng of DNA with cells for 30 minutes then incubate O/N at 37°C

Solutions

YT