SOC Protocol for Transformations

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Transformation Protocol

1. Combine GGA (Golden Gate Assembly) products then redistribute 10 uL of GGA product into PCR tubes.

2. Add GGA product to 50 uL (whole tube) of competent cells.

3. Place tube in ice box for 5 minutes.

4. Meanwhile, put media plate faced down in the bottom shelf of the incubator.

5. Take tube out of ice box after 5 minutes and add 4 volumes of SOC broth (240 mL to 60 mL of cells, 300 mL total) to a 1.5 mL tube, add competent cells in the SOC 1.5 mL tube.

6. Incubate the tubes in a foam rack (sideways in a beaker) in the shaker for 1 hour.

7. After 1 hour, take tubes and spin in centrifuge for 1 minute.

8. Pipette off 250 uL of supernatent and discard.

9. Mix pellet and approximately 50 uL of remaining supernatent.

10. Plate whole (approx. 50 uL) tube on LB + Amp plate.

(if transforming cells in broth, stop after step 6, place all 300 uL of SOC broth and GGA product into the aliquotted tube (8 mL for single, 16 mL for double)