Team:Charlottesville RS/Notebook/material
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Material
Buffers & Solution
Kits
Marker
Enzymes
Plasmidvectors
Synthetic oligonucleotides
Phages
Bacteria
Bacteria Growth Media
Methods
Preparing competent cells
Before lab, bacteria were grown to log (or exponential) phase and 10mL of bacteria were aliquoted into 15 mL sterile plastic tubes. Pelleted cells using tabletop centrifuge at 7000 x g for 10 minutes, or ~4000 rpm. Supernatant was poured off into liquid waste container. Tapped the tube to loosen cell pellet, and pipetted 5 mL of cold CaCL2 solution. Incubated tube on ice for 20 minutes. After incubation, spun cells for 5 minutes at 5000 x g (2500 rpm). Poured off supernatant into liquid waste container. Used pipette to add 1 mL of cold CaCl2 solution and resuspended cells.