1. How does it actually work?

2.  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.  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.  So what actually happens?  Detergents disrupt lipid-lipid interactions compromising cell membrane integrity  Membrane is destroyed or becomes “leaky” depending on lysis buffer

5.  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.  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 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.  A few differences:  Column properties differ: lysing action  Column proprietary; not the traditional silica column