IGEM:Missouri University of Science and Technology/2009/Notebook/Cyanobacteria Mediated Filtration of Coal Flue/2013/09/01

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Date: 09/01/13 People in lab: Emily Puleo, David Pohlman

Title: Making competent cells

Start Time: 4:00 PM

Purpose: Make chemically competent cells

Protocol: 1. Add 1 mL overnight DH5alpha culture to 250 mL SOC, incubate for 2 hrs 2. Chill on ice 10 min 3. Centrifuge 10 min, 3000 rpm 4. Resuspend in 100mL TB buffer (Inoue) 5. Incubate on ice 10 min 6. Centrifuge 10 min 7. Resuspend in 8.67 mL TB with 3.5 mL DMSO 8. Incubate on ice 10 min 9. Aliquot 50 uL per epi tube 10. Store at -80C

Notes: Centrifuge malfunctioned at step 3, cells were not made.

Stop Time: 7:05 PM

Next: Fix centrifuge, try again

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