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Published byPercival Hood Modified over 9 years ago
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CAVEAT!!! Usually we do these procedures with a pure culture ◦ Grow bacteria in broth ◦ Streak on plate ◦ Grow single colony on broth
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Microbial DNA Extraction Bridges 2014
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Cheek Cell
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Bacterial Cell
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Step One: Lysis Breaking apart the cell Physical ◦ Vortex ◦ MicroBeads Chemical ◦ MD1 SDS Disrupts membrane and denatures proteins
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Step Two: Separation from Debris Centrifugation Precipitation of Proteins More Centrifuging Supernatant Pellet
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Step Three: Clean DNA Bind to spin filter ◦ Salt solution ◦ Silica filter Wash with ethanol ◦ Inhibits PCR Elute off of filter
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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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