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Dna Extraction

Dna Extraction. How does it actually work?. Cell Lysis. Lysis buffers are typically detergents such as sodium dodecyl sulfate (SDS) What happens if you add dish soap to a frying pan with water that has a layer of grease on top?. Lysis Cont’d. Another analogy: popping a water balloon

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Dna Extraction

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  1. Dna Extraction How does it actually work?

  2. Cell Lysis • Lysis buffers are typically detergents such as sodium dodecyl sulfate (SDS) • What happens if you add dish soap to a frying pan with water that has a layer of grease on top?

  3. Lysis Cont’d • Another analogy: popping a water balloon • When you pop a balloon, it is like lysing a cell • When the balloon is popped, all of the contents are released

  4. Lysis Cont’d • So what actually happens? • Detergents disrupt lipid-lipid interactions compromising cell membrane integrity • Membrane is destroyed or becomes “leaky” depending on lysis buffer

  5. Wash Stage • The next stage typically involves solutions to remove excess salts and impurities • Additionally, proteases, Rnases, etc. are often added to remove proteins (e.g., histones), RNA, etc.

  6. Wash Cont’d • The wash solution (though separate solutions are sometimes used before) also adjusts pH • At low pH, DNA binds to a column • Historically made of silica—a combination of salts, silica and low pH create conditions that attract DNA

  7. Elution • Elution is a fancy term for release of DNA from column • Water or TE (or other elution buffer) has a neutral or alkiline pH, leading to release of DNA

  8. QIAGEN Kit • A few differences: • Column properties differ: lysing action • Column proprietary; not the traditional silica column

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