cloning

Question 1

what does in vivo mean?

Answer 1

Taking place in a living cell or organism.

Question 2

what does In vitro mean?

Answer 2

Taking place in test tube, flask or petri-dish (a container) outside a living cell or organism.

Question 3

what is the purpose of PCR?

Answer 3

rapidly amplify DNA at an exponential rate

Question 4

what is involved in a typical PCR reaction?

Answer 4

DNA polymerase (Taq); PCR Primers; Template DNA; dNTPs (dATP, dCTP, dGTP, dTTP) and a reaction buffer

Question 5

what will an endonuclease do?

Answer 5

always cut at a specific defined site within the DNA molecule and produce either blunt or sticky DNA ends depending on the nuclease

Question 6

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…

Answer 6

• get the correct orientation of the insert in the plasmid
• prevent the plasmid from re-ligating on itself

Question 7

what’s the conditions in the blue-white selection procedure?

Answer 7

• 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

Question 8

In which order are the steps in a PCR reaction?

Answer 8

Denature -> Anneal -> Extend

Question 9

taq polymerase was first isolated from a bacteria growing…

Answer 9

in a hot spring.