Reading 4

Question 1

What does PCR stand for?

Answer 1

polymerase chain reaction

Question 2

who invented PCR?

Answer 2

Kary Mullis

Question 3

What is needed to carry out a PCR reaction (what do you need to put into the tube for the reaction to occur)?

Answer 3

magnesium

Question 4

Why do you think PCR is such a powerful technique?

Answer 4

Allows rapid amplification of DNA segments, a lot like DNA replication

Question 5

What are the three repeated steps in each cycle of a PCR reaction?

Answer 5

Denaturation , annealing , and extension
(damp aleins explored)

Question 6

What happens during each step?

Answer 6

template into single strands, adding primers, replication of the strands

Question 7

What is the purpose of a primer (in terms of PCR)? How is this similar to a RNA primer that would be synthesized by E. coli primase? How is the primer used in PCR different from that synthesized by E. coli primase?

Answer 7

-starts chain reaction
-dependent on polarity
-primase synthesizes RNA primers while in e coli

Question 8

What is the difference between a deoxyribonucleotide triphosphate (i.e., dATP) and its dideoxyribonucleotide triphosphate (i.e., ddATP)?

Answer 8

-There is no 3`-OH

Question 9

What happens when the polymerase enzyme incorporates a ddNTP during DNA synthesis?

Answer 9

-the synthesis is terminated

Question 10

What is needed to carry out a dideoxy sequencing reaction with radioactively labeled ddNTPs (how many tubes do you need, and what do you need to put into each tube)?

Answer 10

-1 tube
-all 4 ddNTPs with different dyes