Brainscape's Knowledge GenomeTM

Browse over 1 million classes created by top students, professors, publishers, and experts.


See full index

Biochemistry-Nucleic acids > Nucleic Acid Techniques > Flashcards

Nucleic Acid Techniques Flashcards

1
Q

Nucleic Acid Techniques

A
  1. Restriction Mapping
  2. DNA Sequencing
    * Chain Terminator Procedure
    * Automated DNA Sequencing
  3. Recombinant DNA Technology
  4. Polymerase Chain Reaction (PCR)
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q
  1. Restriction Mapping
A
  • Makes use of Restriction enzymes (from bacteria) to digest DNA for analysis of
    fragments generated
    -restriction maps provide framework for locating
  • base sequences
  • genes on small genomes or chromosomes
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

T or F: Bacteria have learned to “restrict” the possibility of attack from foreign DNA by means of “restriction enzymes”

A

True

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

What type of restriction enzymes cleave DNA at selected sites

A

Type II and III

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

Nucleases

A

Enzymes that hydrolyze nucleic acids

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

Endo

A

cleave at an internal location

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

Exo

A

cleave at the end

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

DNase

A

act only on DNA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

RNase:

A

act only on RNA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

Restriction Enzymes 3 types

A

-restriction endonucleases
-cleave dna at internal location
-Isolated chiefly from bacteria
* Type I: requires ATP, non-specific cleavage
* Type II: no ATP requirement, specific cleavage
* Type III: requires ATP, specific cleavage

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

T or F: Cleavage can leave staggered or “sticky” ends or can produce
“blunt” ends

A

true

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

The stability of the DNA double helix is due to

A
  • Hydrogen bonds – between base pairs
  • Electrostatic interactions – mutual repulsion of phosphate groups,
    which makes them most stable on the helix exterior
  • Base-pair stacking interactions
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

intercalating agents

A

Ethidium bromide
* Acridine orange
* Actinomycin D

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

Why can Intercalating Agents Distort the Double Helix

A

Because it is flexible, aromatic macrocycles – flat hydrophobic molecules
composed of fused, heterocyclic rings, can slip between the stacked pairs of bases

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

Ethidium Bromide

A

used to aid in visualization of DNA separated using
gel electrophoresis

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

Acridine Orange

A

used in staining
nucleic acids – helpful in
determining cell cycle

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
17
Q

Actinomycin D

A

an antibiotic shown to have anti-cancer affects. It is shown to have the ability to inhibit transcription by binding DNA at the transcription initiation complex and preventing elongation

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
18
Q

denaturation

A

When heated, DNA double helix will collapse into its two complementary
strands

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
19
Q

hyperchromic effect

A

DNA denaturation results in qualitative changes in the physical properties
* Exposure of bases in denatured DNA results in an increase in absorbance

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
20
Q

Tm

A

he temperature at which half of the DNA is denatured is defined as the
melting temperature

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
21
Q

T OR F: DNA cannot be renatured by cooling

A

False; it can

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
22
Q

reannealing

A

Renaturation requires reassociation of the DNA strands into a
double helix, a process

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
23
Q

Restriction Endonucleases and Gel Analysis..

A

can be used to generate a physical map of genomes, genes, or other segments of DNA

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
24
Q

Restriction maps provide a framework for locating:

A
  • base sequences
  • genes on small genomes or chromosomes.
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
25
Q

restriction–fragment length polymorphisms.

A

Treatment with restriction enzymes yield fragments with different lengths or
what is known

26
Q

2.DNA Sequencing

A

DNA Sequencing used to determine the primary structure of nucleic acids
* Chain Terminator Procedure
* Automated DNA Sequencing

27
Q
  • Primer extension
A

A template DNA base-paired with a
complementary primer is copied by DNA polymerase in the presence of dATP, dCTP, dGTP, dTTP

28
Q

Chain–terminator procedure:

A

1.Fragment of DNA polymerase I (Klenow fragment- polymerase activity)
2.Single Strand of DNA and a DNA primer
3.Nucleoside Triphosphate (dNTPs) – dATP, dGTP, dCTP, dTTP
4.*** small amount of 2’, 3’ dideoxynucleoside triphosphate (ddNTP)

29
Q

ddNTP analog

A

Prevents addition of next nucleotide because 3’-OH needed for phosphodiester bond is replaced with a
3’-H

30
Q

Clone:

A

a collection of molecules or cells, all identical to an original molecule or cell

31
Q

T or F: Gene can be an exact or altered version of natural gene

A

True

32
Q

Plasmids can be cleaved by

A

restriction enzymes, leaving sticky ends

33
Q

How are artificial plasmids constructed?

A

by linking new DNA fragments to the sticky ends of plasmids

34
Q

plasmid

A

naturally occurring, circular, extrachromosomal
DNA molecules found in bacteria

35
Q

Clone + Plasmid –> Vector

A

insertion of foreign DNA into artificial plasmids that serve to carry the foreign DNA insert in E.coli

36
Q

origin of
replication

A

a replicator

37
Q

antibiotic resistance
gene

A

Selectable marker

38
Q

cloning site

A

site where insertion of
foreign DNA will not
disrupt replication or
inactivate essential
markers

39
Q

molecular cloning or genetic engineering

A

The isolation, amplification, and modificiation of
specific DNA sequences

40
Q

Recombinant DNA

A

has 5 basics of cloning

41
Q

Recombinant DNA:
The 5 Basics of Cloning

A
  1. Must be able to cut and rejoin DNA a precise locations
    - Must use sequence specific endonucleases called restriction endonucleases on foreign
    DNA and vector
  2. Select a DNA molecule to serve as a carrier
    - this is know as the vector
  3. Prepare and insert foreign DNA
    - Use DNA ligase to join two pieces of DNA
    - vector with inserted foreign DNA is known as recombinant DNA
  4. Introduce vector/insert into host organism
    - this process is known as transformation
  5. Screen for host cells replicating the hybrid DNA
42
Q

Vector

A

Select a DNA molecule to serve as a carrier

43
Q

Recombinant DNA

A

vector with inserted foreign DNA

44
Q

transformation

A

Introduce vector/insert into host organism

45
Q

1.Plasmids

A

Relatively small
Replicate easily
Carry resistance gene(s) for antibiotics
Contains multiple restriction sites

46
Q

Biologically Functional Chimeric Plasmids : “Transformation”

A

the uptake and replication of exogenous DNA by a recipient cell.To facilitate transformation bacterial cells are rendered permeable to DNA by Ca+

47
Q

An appropriate vector and clone foreign DNA

A
  1. Plasmid
  2. Bacteriophage λ
  3. Bacterial artificial chromosomes (BACs)
48
Q
  1. Bacteriophage
A

produces large amounts of recombnant DNA in easily purified form

49
Q

3.BAC Vectors

A

Bacterial artificial chromosome (BAC) vectors
Allow for cloning of much larger DNA segments
Replicated DNA at a level of one copy per cell

50
Q

Recombinant DNA

A

Detect presence of cloned DNA
1. Antibiotic Resistance
2. Color Change
3. Radioactive Probe

51
Q

Radioactive Probe

A

Identification of a clone with a DNA fragment by hybridization using a radioactive probe

52
Q

PCR( Polymerase Chain Reaction)

A

The small sample of DNA can serve as template for DNA polymerase,
Procedure for amplifying a segment of DNA by
repeated rounds of replication

53
Q

What happens when you don’t have enough DNA for analysis?

A

You make more!! - PCR

54
Q

1 Cycle has 3 temp dependent steps:

A
  1. Denaturation- separation of DNA strands
  2. Annealing- primers anneal to DNA strands
  3. Extension- DNA polymerase synthesizes
    complementary DNA strands
55
Q

matches the sequence of 5’-3’ template strand at the start of the DNA sequence (is complementary and therefore binds the 3’ – 5’ strand) Forward Primer: 5’- ATTAGC -3’

A

Forward primer

56
Q

matches the sequence of 3’-5’ strand at the end of the DNA sequence (is
complementary and therefore binds the 5’ – 3’ strand) Reverse Primer: 5’- CCCGTA -3’

A

Reverse primer

57
Q

In Vitro Mutagenesis

A

(1) Template DNA strands are separated and
amplified by PCR.
(2) Following many cycles of PCR, the DNA
product can be used to transform E. coli cells.
(3) The plasmid DNA can be isolated and
screened for the presence of the unique
restriction site (by restriction endonuclease
cleavage).

58
Q

page 28-31

A
59
Q

Golden rice

A

result of an effort to develop rice varieties that produce provitamin-A
(beta-carotene) as a means of alleviating vitamin A (retinol) deficiencies in the diets of poor
and disadvantaged people in developing countries

60
Q
A

Biochemistry-Nucleic acids (2 decks)

Brainscape helps you reach your goals faster, through stronger study habits.
© 2024 Bold Learning Solutions. Terms and Conditions