This summer we collaborated with the iGEM team at Northwestern University on the InterLab project. They were experiencing difficulties successfully transforming the provided InterLab DNA into their competent DH5a cells. We helped them troubleshoot their issues by providing the protocol we used to transform the DNA. We first rehydrated the provided iGEM DNA from Kit 7 with 10µL sterile water. We then transformed the DNA into JM109 competent cells and miniprepped the colonies to recover the DNA in greater concentrations.

We then used this DNA in our InterLab transformations. The transformation procedure we provided the Northwestern iGEM team is as follows:

1. Add 1µL of the appropriate InterLab DNA into 1.5mL Eppendorf tube containing DH5a competent cells
2. Incubate competent cells on ice for 30 minutes
3. Tap gently every 10 minutes
4. Heat shock DH5a competent cells by placing in 42ºC water bath for 45 seconds
5. For recovery, place cells on ice for 2 minutes
6. Add 300µL of SOC media to rescue DH5a competent cells
7. Place cells in 37ºC incubator for 1 hour
8. Spin cells in microcentrifuge at 5K rpm for 2 minutes
9. Remove the upper 200µL of supernatant fluid
10. Resuspend cells in the remaining 100µL of SOC media
11. Plate 50µL of DH5a competent cells with InterLab DNA on LB + CAM plates
12. Incubate transformed DH5a competent cells in 37ºC incubator for 16 hours

Using the procedure we provided, the Northwestern iGEM team was able to successfully transform their cells and complete the InterLab study.