miniprep lecture

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.

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

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

Protocol

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

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

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

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

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.