Dna Extraction

DNA EXTRACTIONHow 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 When you pop a

balloon, it is like lysing a cell

When the balloon is popped, all of the contents are released

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

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.

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

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

QIAGEN Kit A few differences:

Column properties differ: lysing action

Column proprietary; not the traditional silica column