Miniprep Lecture
8
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.
Transcript of 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 endup working in a lab then you likely will have done orwill 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.
