IGEM:Missouri University of Science and Technology/2009/Notebook/Cyanobacteria Mediated Filtration of Coal Flue/2013/09/23

Cyanobacteria Mediated Filtration of Coal Flue Gas

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Date: 09/23/13 People in lab: Kelsey Crossen, Emily Puleo Title: Making chemically competent cells Start Time: 3:05 PM Purpose: To make chemically competent cells Protocol: 1. Add overnight culture to 50 mL SOB, incubate 2 hrs 2. Chill on ice 10 min 3. Centrifuge 10 min 4500 rpm at 4C, decant supernatant 4. Resuspend in 5mL TB buffer (Inoue) 5. Incubate on ice 10 min 6.Centrifuge 10 mins, 4500 rpm, 4C 7. Resuspend in 930 uL TB and 70 uL DMSO 8. Incubate on ice 10 mins 9. Aliquot 50uL per epi tube 10. Store in -80C freezer Products: Sample Label Description 9/23/13 Comp cells Chemically competent DH5alpha Stop Time: 4:38 PM