Part D: Lecture 35

Question 1

DNA replication requires____

Answer 1

primer
monomer
ssDNA template

Question 2

dsDNA denatures when ____ and renature ____

Answer 2

heated
cooled

Question 3

during renaturation ______ of ssDNA can pair

Answer 3

any complementary pieces

Question 4

oligonucleotides are ____ that are ___ long

Answer 4

synthetic ssDNA
20 nt

Question 5

oligos bind to ___ and serve as ____

Answer 5

complementary ssDNA
primers for invitro DNA replication

Question 6

purpose of PCR?

Answer 6

in vitro exponential amplification of a small region of DNA

Question 7

limiting factors of PCR

Answer 7

1) must know the sequence at both ends of the region to be amplified
2) region amplified can’t be > 5kb

Question 8

cycle from PCR

Answer 8

1)95 C: denaturation
2)55 C: primers anneal
3)72 C: extension of DNA

Question 9

PCR machine is a ___

Answer 9

thermocycler machine (changes temperature)

Question 10

PCR reaction

Answer 10

template DNA
Taq DNA Pol
left/forward oligo primers
right/reverse oligo primers
dNTPs

Question 11

Entire PCR reaction has ___ cycles

Answer 11

30

Question 12

each cycle of PCR, the products become ___

Answer 12

templates for next cycle

Question 13

after cycling, there is a temperature hold at?

Answer 13

4 C

Question 14

after 30 cycles, PCR tube contains ___

Answer 14

original template DNA
some unused dNTPs + primers
few variable length DNA
lots of constant length DNA = < 1 billion; seen on gel

Question 15

primers become _____

Answer 15

part of the PCR product

Question 16

Primers are where in PCR products?

Answer 16

5’ end of both strands

Question 17

PCR primers are where in PCR template?

Answer 17

on opposite strands

Question 18

uses of PCR

Answer 18

1)amplify DNA for sequencing
2)detect difference in the length of a gene (mutant alleles)

Question 19

hemizygous

Answer 19

individual who has only one member of a chromosome pair or chromosome segment rather than the usual two (usually X-linked)