Ch. 19 Part 1

Question 1

What is PCR

Answer 1

DNA amplification in a test tube; used to make a lot of a target sequence of DNA

Question 2

What are the components of PCR

Answer 2

Template DNA, Taq polymerase, dNTPs, primers (forward and reverse), buffers

Question 3

Primers are important because

Answer 3

they are needed to start DNA synthesis, and they define the outer bounds of your target sequence

Question 4

What are the 3 stages of PCR

Answer 4

1- denature
2- annealing
3- elongation

Question 5

True or false: the primers used in PCR are made of RNA

Answer 5

False: made of DNA

Question 6

What does gel electrophoresis do?

Answer 6

it separates molecules based on their size; nucleic acids will travel towards a positive pole

Question 7

Do smaller or larger DNA fragments run farther in gel electrophoresis?

Answer 7

smaller

Question 8

Where do most restriction enzymes cut?

Answer 8

a palindromic sequence

Question 9

How can restriction enzymes be used in recombinant DNA?

Answer 9

cutting different sources of DNA with the same sticky-end restriction enzyme allows them to be recombined

Question 10

What is a vector?

Answer 10

an agent that can carry DNA into a cell or organism

Question 11

How to make lots of a sequence with a cloning vector in plasmid cloning

Answer 11

cut the DNA you want to clone and the plasmid with the same sticky-end restriction enzyme; you mix the cut sample DNA and cut plamid together with DNA ligase; and once the target sequence is in the plasmid, the plasmid is introduced to bacteria through transformation

Question 12

What are the 3 essential components of a plasmid vector

Answer 12

multiple cloning site, ori, and the selectable marker (there is also a screenable marker)

Question 13

In a plamid vector: the ori

Answer 13

ensured that the vector is replicated within the cell

Question 14

In a plamid vector: the MCS

Answer 14

is where your target sequence goes

Question 15

In a plamid vector: the selectable marker

Answer 15

enables any cells containing the vector to be selected/ identified

Question 16

transformation (plasmid vectors)

Answer 16

when the cells take up the recombinant plasmid (not all cells take up plamid)

Question 17

Why do scientists use the blue white screening in plasmid cloning

Answer 17

to screen for bacteria carrying the recombinant plasmid

Question 18

Which of these is not a component of the plasmid cloning vector: Ori, MCS, antibiotic resistant gene, essential genes for structure and metabolism, or none of the above?

Answer 18

essential genes for structure and metabolism

Question 19

If no foreign DNA has been inserted into the partial lacZ gene then…

Answer 19

the cell will produce B- galactosidase (shows up blue)

Question 20

If foreign DNA has been successfully added…

Answer 20

it disrupts the front end of the lacZ gene and B-galactosidase is not produced (white)

Question 21

expression vectors

Answer 21

used when the goal is not just to replicate the gene- but to also produce the protein it encodes; it contains sequences required for transcription and translation in bacterial cells

Question 22

the operon sequence in the expression vector is responsible for what

Answer 22

allows inserted DNA to be transcribed and translated

Question 23

What is another important part of an expression vector besides the operon sequences?

Answer 23

sequences that regulate the desired gene (turn the gene on or off)