Bio 006 - Lab 5: Plasmid Mini-Prep

Plasmid Mini-Prep

Process of extracting plasmid DNA from bacteria.

Transformation

Inserting plasmid DNA into bacteria.

Centrifuge

Machine that separates materials by density.

Supernatant

Less-dense liquid layer after centrifugation.

Pellet

Higher-density solids at the bottom after centrifugation.

GTE Buffer

Reagent used to destabilize cell membranes.

Sodium Dodecyl Sulfate

breaks down cell membranes (aka SDS)

Sodium Hydroxide

denatures genomic DNA.

KOAc

Helps renature plasmid DNA after lysis.

Isopropanol

Precipitates DNA from the supernatant.

Ethanol

Washes plasmid DNA during purification.

TE Buffer

Protects DNA for subsequent experiments.

Minicentrifuge

Smaller centrifuge for 6 or fewer samples.

Microcentrifuge

Larger centrifuge; holds many samples, more sensitive.

Vortex

Device used to mix solutions rapidly.

Inverting

Technique for mixing by turning tubes upside down.

Finger Vortexing

Mixing technique using fingers to agitate solution by flicking

Wicking

Technique to absorb liquid using absorbent material.

Lysis

Process of breaking down cell membranes.

Purification

Isolating plasmid DNA from other cellular components.

Recombinant Plasmid

Plasmid containing DNA from different sources.