Lab 7 Transformation Flashcards
Refers to the uptake of foreign DNA (such as a plasmid) by a cell. Introducing plasmid into bacteria.
Transformation
A small circular DNA molecule found in some bacteria that contains an origin of replication.
Plasmid
A plasmid used to clone a foreign gene
Cloning vector
In the lab when transforming bacteria with plasmid, very very few cells actually take up the plasmid
Therefore, a plasmid typically Carries a gene for antibiotic resistance so that transformed bacteria can grow in the presence of antibiotic.
To be used as a vector for recombinant DNA, a plasmid much have…
- An origin of replication (ori)
- A selectable marker (antibiotic resistance gene is typically used)
- A convenient way to insert the recombinant DNA, usually a multiple cloning site (MCS)
(MCS) Multiple cloning site
An area of the plasmid that has a lot of restriction sites.
A restriction enzyme usually makes many cuts yielding
Restriction fragments
Cut DNA molecules at specific DNA sequences called restriction sites.
Bacterial restriction enzymes
What growth was expected on the plate with +DNA on LB (growth medium)
Lots of bacteria(ecoli) , ALL the bacteria plated could grow in absence of ampicillin.
What growth was expected on the plate with -DNA on LB (growth medium)
With or without the plasmid DNA , ALL the bacteria that was plated could grow in the absence of ampicillin.
What growth would be expected on the plate with -DNA on LB + amp
No growth, without the plasmid DNA, which contained the amp gene, non of the bacteria found grow in the presence of ampicillin
What growth was expected on the plate with +DNA on LB + amp
Colonies. Each colony grows from a single bacterium that was transformed with the plasmid DNA and could grow in the presence of ampicillin
Plasmid
A small DNA molecule within bacterial cell
