Unit 4.5

Question 1

Technology used to amplify DNA

Answer 1

PCR

Question 2

PCR

Answer 2

Polymerase Chain Reaction

Question 3

The production of multiple copies of a sequence of DNA

Answer 3

Amplifying

Question 4

Components of PCR (6)

Answer 4

1. DNA sample
2. Primers
3. Nucleotides
4. Taq Polymerase
5. Mix Buffer
6. PCR Tube

Question 5

PCR Steps: DAER (4)

Answer 5

1. Denature
2. Anneal Primers
3. Extend
4. Repeat

Question 6

A special type of polymerase found in extreme environments like hot springs that is resistant to heat

Answer 6

Taq polymerase

Question 7

PCR steps (DAER) but specific (4)

Answer 7

1. D: heat is used to denature the enzymes holding the DNA strands together
2. A: the strands are cooled and DNA primers are attached
3. E: DNA is synthesized along the strands by taq polymerase (may be warmer than during annealing)
4. R: the cycle is repeated until enough strands are created

Question 8

Uses of PCR (7)

Answer 8

1. Duplicating DNA at a crime scene for experimentation
2. Diagnosing diseases
3. Screening for mutations
4. Organism classification
5. Genotyping
6. Gene therapy
7. Paternity tests

Question 9

Enzyme used in reverse transcription PCR to convert RNA to DNA

Answer 9

Reverse transcriptase

Question 10

What temp is DNA denatured at for PCR

Answer 10

95C

Question 11

What temp are primers annealed to DNA in PCR

Answer 11

60C

Question 12

What temp does taq polymerase synthesizes nucleotides in PCR

Answer 12

72C

Question 13

Average number of cycles of PCR so the DNA can be visualized

Answer 13

25-35

Question 14

Equation for number of DNA copies made during PCR

Answer 14

2^n (n = number of cycles of PCR)

Question 15

PCR test for COVID testing

Answer 15

Real-Time Reverse Transcription Polymerase Chain Reaction test (rRT-PCR test)

Question 16

Inventor of the PCR technique

Answer 16

Dr. Kary Mullis

Question 17

Bacteria produce these to break down phage DNA and prevent infection

Answer 17

Restriction Enzymes (RE)

Question 18

Other word for restriction enzymes

Answer 18

Restriction endonucleases

Question 19

Each RE cuts a specific ___ DNA sequence that is found in the viral DNA of an infecting ___

Answer 19

Palindromic, phage

Question 20

A virus that replicates within bacteria

Answer 20

Phage

Question 21

Cutting viral DNA prevents the viral ___ from being used to make more virus

Answer 21

Genome

Question 22

Uses of REs (3)

Answer 22

1. gene cloning
2. research
3. DNA testing

Question 23

REs “cut” both strands of the DNA by breaking the ___ bonds between the nucleotides, called a ___ cut

Answer 23

Covalent, double-stranded

Question 24

DNA fragments from REs can be put into ___ or ___ and used to make bacteria produce the gene product of that DNA sequence through ___

Answer 24

Vectors, plasmids, bacterial transformation

Question 25

Lab protocol that causes bacteria to take in and produce proteins from non-bacterial DNA found in vectors/plasmids

Answer 25

Bacterial transformation

Question 26

Steps of bacterial transformation (4)

Answer 26

1. Incubate vector/plasmid with bacteria that have been heat shocked to open up pores in their membrane (vector/plasmid will have an antibiotic resistance gene in it) 2. Bacteria take up the plasmids with your gene of interest in it 3. Grow colonies of these bacteria on nutrient media that contain antibiotics (only bacteria with the plasmid will survive) 4. Bacteria use their own protein synthesis machinery to make the gene product of your gene (this is how they make insulin for diabetes patients)

Question 27

How to open up pores in a vector/plasmid membrane

Answer 27

Heat shocking

Question 28

Vectors/plasmids have ___ resistance genes

Answer 28

Antibiotic

Question 29

single bases in genes that differ based on the allele someone has for a specific gene

Answer 29

Small Nucleotide Polymorphisms (SNPs)

Question 30

Gel used for DNA fingerprinting/gel electrophoresis

Answer 30

Agarose gel

Question 31

A technique used to separate DNA fragments based on size and charge by pulling them through a gel using an electric current

Answer 31

Gel electrophoresis

Question 32

DNA fragments are __ charged, so they move towards the ___ electrode in gel electrophoresis

Answer 32

Negatively, positively

Question 33

Because all DNA fragments have the same amount of charge per mass, ___ fragments move through gel faster than ___ ones

Answer 33

Small, large

Question 34

A sequence in DNA where two or more sequential base pairs are repeated. Useful for DNA profiling

Answer 34

Tandem repeats

Question 35

Features of DNA fragments used to separate them during gel electrophoresis (2)

Answer 35

1. Size 2. Charge

Question 36

What is agarose

Answer 36

A polysaccharide polymer

Question 37

What is agarose usually made from

Answer 37

Seaweed