Laygo, Demi Jamie Q.

2014 48931

THQ MBB1
DNA Extraction

1. NaCl
Sodium chlorides role is to remove proteins that are bound to the
DNA and to help keep proteins dissolved in the aqueous layer so
they dont precipitate in the alcohol along with the DNA.
2. Baking Soda
Baking sodas role is to serve as a buffering system that raises the
pH (near 8) and to release the DNA from bound proteins since
DNA is only soluble at a pH near physiological levels.
3. Detergent
Detergents role is to help dissolve the cell membrane, which is a
lipid bilayer.
4. Hot water bath
Hot water baths role is to soften the phospholipids in the cell
membrane and to denature the DNAse enzymes. These enzymes
would cut the DNA into fragments and would make it too small for
extraction if it werent for the heat treatment.
5. Cold water bath
Cold water baths role is to slowdown the breakdown of DNA. If
not done immediately after the hot water bath, the DNA will be
fragmented and will not spool.
6. Ethanol
Ethanols role is to separate the DNA from other macromolecules
and cell contents. DNA is not soluble in alcohol thus the DNA
present at the water-alcohol interface precipitates out of solution,
allowing it to be extracted.
References:
https://biology.mit.edu/sites/default/files/DNASpooling.pdf
http://dwb4.unl.edu/Chem/CHEM869N/CHEM869NLinks/www.framingham.k12.ma.us/fhs/dept
s/science/Biotechnology/protocols.html
http://www.gs.washington.edu/outreach/dhillon_dnaprocedure.pdf