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Gr. 12 ap bio > Unit 2 Pt2 Test > Flashcards

Unit 2 Pt2 Test Flashcards

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1
Q

VNTR

A

Variable Number Tandem Repeats

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2
Q

PCR

A

Polymerase Chain Reaction

Synthetic method (in vitro) to produce copies of DNA

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3
Q

Isolating DNA (3)

A
  • cell membranes are disrupted
  • DNA precipitation (ethanol used to dehydrate and aggregate DNA)
  • DNA isolation/storage
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4
Q

Restriction endonucleases

A

Enzymes that cut DNA backbones at specific sequences through hydrolysis

Naming: 
Genus (1 letter) 
Species (2 letters) 
Strain (1 letter) 
# enzyme in strain
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5
Q

Recognition site

A

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

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6
Q

Used to chemically join two pieces of DNA

A

T4 DNA ligase

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7
Q

Recombinant DNA

A

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

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8
Q

Gene cloning

A

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

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9
Q

Bacteria contain circular DNA as:

A
  1. A requirement (chromosome)

2. Extra material (plasmids)

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10
Q

Gene cloning (3)

A

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

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11
Q

Methods to help bacteria pick up plasmid:

A
  1. Heat-shocking (leaky, permeable)

2. Electroporation (permeable)

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12
Q

Who developed PCR

A

Kary Mullis

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13
Q

Minimum requirements for DNA polymerase:

A
  1. Template strand
  2. Primers
  3. dNTPs
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14
Q

Steps for PCR cycle (3):

A
  1. DNA Strand denaturation (95°C)
  2. Primer annealing (50-65°C)
  3. DNA Strand synthesis (72°C)
    Taq polymerase
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15
Q

PCR applications

A

Genetic screening

Forensic analysis

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16
Q

Target sequence is achieved after how many cycles?

A

3

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17
Q

Gel electrophoresis

A

Moving DNA through a gel medium using an electric current

18
Q

Components of gel electrophoresis setup:

A
  • well
  • buffer (not water)
  • samples (DNA)
  • electrodes
  • frame
19
Q

Short DNA & Long DNA:

A

Short DNA:
•moves through gel easily
•farther

Long DNA:
•moves w more effort
•closer

20
Q

Gel medium made from:

A

Agarose (seaweed extract)

Polyacrylamide (artificial polymer, higher resolution)

21
Q

Resolution

A

How well separated the DNA pieces are/need to be

22
Q

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

A 
Ethidium bromide (untwists the DNA a little)
UV light box
23
Q

DNA ladder

A

Aka DNA marker

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

24
Q

RFLP

A

Restriction Fragment Length Polymorphism

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

25
Q

Large genomes cut with restriction enzymes look like ____ after electrophoresis

A

Smear

26
Q

Southern blotting

A

Transfer of gel electrophoresis DNA to a nylon membrane (no electricity)

Stacked:
Weight
Absorbent paper
Nylon membrane
Gel
(DNA moves up)
27
Q

RFLP probe

A

Specific, radioactive DNA sequences are exposed to the membrane

28
Q

RFLP analysis

A

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

29
Q

Photographic film used to detect the (radioactive) sequence

A

RFLP autoradiograph

Compared with blots to see whose DNA lit up

30
Q

Who invented the DNA sequencing method?

A

Fredrick Sanger

31
Q

Fredrick Sanger developed the ______

A

Dideoxy termination sequencing method

32
Q

DNA is sequencing is based on ___

A

How DNA is replicated

33
Q

The sequence of bases could be determined if

A
  1. DNA polymerase was stopped at each base

2. The last nucleotide added could be identified (AGCT)

34
Q

Components of a sequencing reaction:

A
  1. DNA polymerase
  2. DNA template strand
  3. Primer
  4. Deoxyribonucleotides (dNTPs)
  5. Dideoxyribonucleotides (ddNTPs)
35
Q

What does dideoxyribonucleotide lack that stops the elongation?

A

3’ OH group

36
Q

What % for ddNTPs

A

10%

37
Q

What gel medium is suited for dideoxy termination sequencing

A

Polyacrylamide

38
Q

What is microarray used for?

A

Examine thousands of mRNA strands all at the same time

39
Q

What are the two microarray samples?

A
  1. Control (wild/normal)

2. Treatment/Abnormal

40
Q

Steps to Microarray Analysis (4)

A
  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
41
Q

Focus on green/red dots

A

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

(mRNA production)

42
Q

List all dNTPs (4)

A

Deoxyadenosine triphosphate
Deoxyguanosine triphosphate
Deoxythymidine triphosphate
Deoxycytidine triphosphate

Gr. 12 ap bio (8 decks)

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