Miniprep Lecture

1. Minipreps
The term “miniprep” refers to a procedure or
protocol used to purify plasmid DNA.
If you have worked in a lab previously or if you end
up working in a lab then you likely will have done or
will do many, many minipreps.

2. Nucleic Acids Research (1979), 7: 1513 - 1523

3. http://tools.invitrogen.com/content/sfs/manuals/purelink%20_quick_plasmid_qrc.pdf

4. Protocol
http://tools.invitrogen.com/content/sfs/manuals/purelink%20_quick_plasmid_qrc.pdf

5. http://tools.invitrogen.com/content/sfs/manuals/purelink%20_quick_plasmid_qrc.pdf

6. http://tools.invitrogen.com/content/sfs/manuals/purelink%20_quick_plasmid_qrc.pdf

7. One of the solutions (e.g., the Precipitation Buffer)
contained a chaotropic agent or salt.
What the heck is a chaotropic agent?
“A chaotropic agent', also known as chaotropic
reagent and chaotrope, is a substance which disrupts
the three dimensional structure in macromolecules
such as proteins, DNA, or RNA. Chaotropic agents
interfere with stabilizing intramolecular interactions
mediated by noncovalent forces such as hydrogen
bonds, Van der Waals forces, and hydrophobic effects.”
http://en.wikipedia.org/wiki/Chaotropic_agent

8. In the presence of a suitable chaotropic reagent,
DNA will bind to siliceous materials.
DNA remains bound to the resin when EtOH is
used to rinse the resin because EtOH by itself
will precipitate DNA.
A final rinse in dH2O causes the DNA to elute
from the resin.