cloning Flashcards
what does in vivo mean?
Taking place in a living cell or organism.
what does In vitro mean?
Taking place in test tube, flask or petri-dish (a container) outside a living cell or organism.
what is the purpose of PCR?
rapidly amplify DNA at an exponential rate
what is involved in a typical PCR reaction?
DNA polymerase (Taq); PCR Primers; Template DNA; dNTPs (dATP, dCTP, dGTP, dTTP) and a reaction buffer
what will an endonuclease do?
always cut at a specific defined site within the DNA molecule and produce either blunt or sticky DNA ends depending on the nuclease
Cutting the plasmid, and the DNA to be inserted, with two different restriction enzymes is better than cutting with one because cutting with two helps to…
- get the correct orientation of the insert in the plasmid
- prevent the plasmid from re-ligating on itself
what’s the conditions in the blue-white selection procedure?
- All the blue colonies on the plate contain the plasmid and the plasmid does not contain the insert
- All the colonies on the plate contain the plasmid.
- All the white colonies on the plate contain the plasmid and the plasmid contains the insert
In which order are the steps in a PCR reaction?
Denature -> Anneal -> Extend
taq polymerase was first isolated from a bacteria growing…
in a hot spring.
