1 / 8

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

ossie
Télécharger la présentation

Dna Extraction

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  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 alkaline pH, leading to release of DNA

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

More Related