Unit 2 Pt2 Test

Question 1

VNTR

Answer 1

Variable Number Tandem Repeats

Question 2

PCR

Answer 2

Polymerase Chain Reaction

Synthetic method (in vitro) to produce copies of DNA

Question 3

Isolating DNA (3)

Answer 3

• cell membranes are disrupted
• DNA precipitation (ethanol used to dehydrate and aggregate DNA)
• DNA isolation/storage

Question 4

Restriction endonucleases

Answer 4

Enzymes that cut DNA backbones at specific sequences through hydrolysis

Naming: 
Genus (1 letter) 
Species (2 letters) 
Strain (1 letter) 
# enzyme in strain

Question 5

Recognition site

Answer 5

The DNA sequence which restriction enzymes bind to
(Usually 4-8bp)
PALINDROMIC

Question 6

Used to chemically join two pieces of DNA

Answer 6

T4 DNA ligase

Question 7

Recombinant DNA

Answer 7

Complementary sticky ends from different pieces of DNA can be joined together

Question 8

Gene cloning

Answer 8

Method of inserting new DNA into simple organism to make many copies of the gene and protein

Used to examine function and structure of gene + products

Question 9

Bacteria contain circular DNA as:

Answer 9

1. A requirement (chromosome)

2. Extra material (plasmids)

Question 10

Gene cloning (3)

Answer 10

a) Creation of recombinant DNA plasmid
b) bacterial transformation
c) screening for successful clones

Question 11

Methods to help bacteria pick up plasmid:

Answer 11

1. Heat-shocking (leaky, permeable)

2. Electroporation (permeable)

Question 12

Who developed PCR

Answer 12

Kary Mullis

Question 13

Minimum requirements for DNA polymerase:

Answer 13

1. Template strand
2. Primers
3. dNTPs

Question 14

Steps for PCR cycle (3):

Answer 14

1. DNA Strand denaturation (95°C)
2. Primer annealing (50-65°C)
3. DNA Strand synthesis (72°C)
   Taq polymerase

Question 15

PCR applications

Answer 15

Genetic screening

Forensic analysis

Question 16

Target sequence is achieved after how many cycles?

Answer 16

3

Question 17

Gel electrophoresis

Answer 17

Moving DNA through a gel medium using an electric current

Question 18

Components of gel electrophoresis setup:

Answer 18

• well
• buffer (not water)
• samples (DNA)
• electrodes
• frame

Question 19

Short DNA & Long DNA:

Answer 19

Short DNA:
•moves through gel easily
•farther

Long DNA:
•moves w more effort
•closer

Question 20

Gel medium made from:

Answer 20

Agarose (seaweed extract)

Polyacrylamide (artificial polymer, higher resolution)

Question 21

Resolution

Answer 21

How well separated the DNA pieces are/need to be

Question 22

DNA Is strained w ____ and a ___________ to see fluorescent DNA bands

Answer 22

Ethidium bromide (untwists the DNA a little)
UV light box

Question 23

DNA ladder

Answer 23

Aka DNA marker

DNA sample with known bp’s. Used as reference/guide

Question 24

RFLP

Answer 24

Restriction Fragment Length Polymorphism

Polymorphism - differences between DNA sequences (individuals can be differentiated by slight changes)

Question 25

Large genomes cut with restriction enzymes look like ____ after electrophoresis

Answer 25

Smear

Question 26

Southern blotting

Answer 26

Transfer of gel electrophoresis DNA to a nylon membrane (no electricity) ``` Stacked: Weight Absorbent paper Nylon membrane Gel (DNA moves up) ```

Question 27

RFLP probe

Answer 27

Specific, radioactive DNA sequences are exposed to the membrane

Question 28

RFLP analysis

Answer 28

Membrane DNA is denatured to expose target sequences for the DNA probe Radioactive DNA only binds to membrane where the sequences are a complementary match

Question 29

Photographic film used to detect the (radioactive) sequence

Answer 29

RFLP autoradiograph | Compared with blots to see whose DNA lit up

Question 30

Who invented the DNA sequencing method?

Answer 30

Fredrick Sanger

Question 31

Fredrick Sanger developed the ______

Answer 31

Dideoxy termination sequencing method

Question 32

DNA is sequencing is based on ___

Answer 32

How DNA is replicated

Question 33

The sequence of bases could be determined if

Answer 33

1. DNA polymerase was stopped at each base | 2. The last nucleotide added could be identified (AGCT)

Question 34

Components of a sequencing reaction:

Answer 34

1. DNA polymerase 2. DNA template strand 3. Primer 4. Deoxyribonucleotides (dNTPs) 5. Dideoxyribonucleotides (ddNTPs)

Question 35

What does dideoxyribonucleotide lack that stops the elongation?

Answer 35

3’ OH group

Question 36

What % for ddNTPs

Answer 36

10%

Question 37

What gel medium is suited for dideoxy termination sequencing

Answer 37

Polyacrylamide

Question 38

What is microarray used for?

Answer 38

Examine thousands of mRNA strands all at the same time

Question 39

What are the two microarray samples?

Answer 39

1. Control (wild/normal) | 2. Treatment/Abnormal

Question 40

Steps to Microarray Analysis (4)

Answer 40

1. mRNA is isolated from tissue 2. cDNA is made using mRNA as template (labelled with a fluorescent protein) 3. Fluorescent cDNA is place into the microarray 4. Laser scanner to locate fluorescence

Question 41

Focus on green/red dots

Answer 41

These represent mRNA that are different between the control and the test sample (mRNA production)

Question 42

List all dNTPs (4)

Answer 42

Deoxyadenosine triphosphate Deoxyguanosine triphosphate Deoxythymidine triphosphate Deoxycytidine triphosphate