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CAVEAT!!!

CAVEAT!!!. Usually we do these procedures with a pure culture Grow bacteria in broth Streak on plate Grow single colony on broth. Microbial DNA Extraction. Bridges 2014. Cheek Cell. Bacterial Cell. Step One: Lysis. Breaking apart the cell Physical Vortex MicroBeads Chemical MD1 SDS

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CAVEAT!!!

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  1. CAVEAT!!! • Usually we do these procedures with a pure culture • Grow bacteria in broth • Streak on plate • Grow single colony on broth

  2. Microbial DNA Extraction Bridges 2014

  3. Cheek Cell

  4. Bacterial Cell

  5. Step One: Lysis • Breaking apart the cell • Physical • Vortex • MicroBeads • Chemical • MD1 • SDS • Disrupts membrane and denatures proteins

  6. Step Two: Separation from Debris • Centrifugation • Precipitation of Proteins • More Centrifuging Supernatant Pellet

  7. Step Three: Clean DNA • Bind to spin filter • Salt solution • Silica filter • Wash with ethanol • Inhibits PCR • Elute off of filter

  8. Contamination • It is important when working with bacteria to use sterile technique • Wipe work area, pipets, and pipet boxes with EtOH • Use aerosol filter pipets • Keep tips covered • Change tips • Especially if potentially contaminated • Change gloves often! • Keep tubes closed as much as possible • Do NOT touch inside of tubes • STAY ORGANIZED!

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