Fundamentals of PCR

Question 1

Which of the following are the main steps of PCR:
I. Denaturation
II. Extension
III. Annealing
IV. Reversion

a. I, II, IV
b. I, II, III, IV
c. I and III only
d. I, II, III

Answer 1

D

Question 2

step in PCR wherein the DNA is subjected at 95°C to separate the strands

Answer 2

denaturation

Question 3

step in PCR wherein the primers are subjected at 40°C-65°C to bind with their complementary sequences

Answer 3

annealing

Question 4

at 72°C DNA polymerase is subjected to add nucleotides to the 3’ ends of the primers

Answer 4

extension

Question 5

what are the building materials of the DNA polymerase

Answer 5

dNTPS

Question 6

what is the basis of the PCR

Answer 6

temperature change

Question 7

what makes the unwinding of the double-stranded DNA possible

Answer 7

helicase

Question 8

what is the temperature needed in denaturation of DNA in PCR

Answer 8

95°C

Question 9

what is the temperature needed in annealing primers in PCR

Answer 9

40°C-65°C

Question 10

what is the temperature needed in extending the DNA chain in PCR

Answer 10

72°C

Question 11

these are the heat-stable DNA polymerase used from hot springs bacterium

Answer 11

Taq DNA polymerase

Question 12

what bacteria does the Taq DNA polymerase purified from

Answer 12

Thermus aquaticus

Question 13

what temperature is the maximal enzymatic activity of Taq polymerase

Answer 13

75°C to 80°C

Question 14

This occurs at 95°C mimicking
the function of helicase in the
cell.

Answer 14

denaturation

Question 15

Primers bind to the
complimentary sequence on the
target DNA.

Answer 15

annealing/binding

Question 16

These are chosen
such that one is complimentary
to the one strand at one end of
the target sequence and that
the other is complimentary to
the other strand at the other end
of the target sequence

Answer 16

primers

Question 17

two types of primers in PCR

Answer 17

reverse and forward

Question 18

true or false: The
DNA is antiparallel. The first
strand runs on 5’ to 3’ and the
other runs on 5’ to 3’ direction.

Answer 18

False. The first strand runs on 5’ to 3’ and the other runs on 3’ to 5’ direction.

Question 19

DNA polymerase catalyzes the
extension of the strand in the
5-3 direction, starting at the
primers, attaching the
appropriate nucleotides (A-T,
C-G)

Answer 19

Extension/Primer Extension

Question 20

true or false: The PCR reaction will amplify the DNA section between the two primers

Answer 20

true

Question 21

true or false: If the DNA sequence is known, primers can be developed to amplify any piece of an organism’s DNA

Answer 21

true

Question 22

the DNA of interest is amplified by a power of ____ for each PCR cycle

Answer 22

2

Question 23

This uses repeated cycles of
heating and cooling to make
many copies of a specific region
of DNA

Answer 23

polymerase chain reaction (PCR)

Question 24

what enzyme binds to the prime sequences and adds nucleotides to extend the DNA chain

Answer 24

Taq polymerase