Chemical Transformation
Before starting
- Heat hot plate or water bath to 42°C
- Optional: Warm selection plates to 37°C
Transformation
- Thaw chemically competent cells on ice for 10-15 minutes
- Add 40ul cells to fresh 1.7mL tube
- Strongly recommended: set up an extra sample as a negative control. Do not add DNA to this sample
- Add DNA
- If using a ligation product add up to 10ul of sample
- If using supercoiled plasmid add 100ng
- Incubate on ice for 30 minutes
- Heat shock cells on hot plate (or water bath) for 30-45s* @ 42°C
- Incubate on ice for 2-5 minutes
- Recovery:
- Option 1: Recover in 200ul
- Add 200 μL SOC and shake gently for 1-2 hours @ 37°C
- Option 2: Recover in 960ul
- Add 960ul of SOC and shake gently for 1-2 hours @ 37°C
- Gently spin and remove supernatant
- Resuspend pellet in 200ul LB
- Plate 100-200ul cells onto selection plates
- If high efficiency is expected, we suggest plating a lower volume
- If the expected efficiency is unknown, we suggest also plating a 1:10 dilution
- Once dry, turn upside down (agar on top) and incubate overnight @ 37°C
* The optimal time depends on the cells
SOC (1L)
| Tryptone | 20g |
| Yeast Extract | 5g |
| MgSO4 | 4.8g |
| Dextrose | 3.6g |
| NaCl | 0.5g |
| KCl | 0.186g |
