Brainscape's Knowledge GenomeTM

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


See full index

Genetics 1 > Topic 10: DNA Technology and Applications in Genetics > Flashcards

Topic 10: DNA Technology and Applications in Genetics Flashcards

You may prefer our related Brainscape-certified flashcards:
Biology 101 flashcards
1
Q

what are the three new technologies for molecular genetics?

A

polymerase chain reaction (PCR)
DNA sequencing (snager method, next generation, third generation)
Restriction enzymes

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

who developed PCR in 1983?

A

Kary Mullis

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

what are the three main steps of PCR?

A
  1. DNA denaturation (1-2 mins)
  2. Primer annealing (1-2 mins)
  3. Polymerase extension (3-5 mins)
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

to what temperature do you heat the reaction mixture to denature DNA in PCR?

A

95 degrees Celcius

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

PCR produces DNA fragments of varying lengths, how can we visualize these fragments?

A

gel electrophoresis

separates DNA fragments by their base pair lengths, longer fragments are heavier and move slower than smaller fragments

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

di-deoxynucleotides are like dNTPs but lack two oxygen, what is this then abbreviated to be?

A

ddNTPs

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

what is the official scientific name for the “Sanger method of DNA sequencing?

A

dideoxynucleotide DNA sequencing

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

true/false: lack of OH prevents the ddNTP from forming phosphodiester bonds

A

true

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

true/false: in dideoxynucleotide DNA sequencing, 5-3’ polarity corresponds to the smaller-to-larger fragment length direction

A

true

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

how does next-generation DNA sequencing (NGS) work? why is it faster and cheaper than the Sanger method

A

uses computational power to put fragments together into genome-wide consensus sequence

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

true/false: third-generation DNA sequencing (TGS) isn’t very similar and parallel to next-generation DNA sequencing

A

false, its very similar and parallel

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

restriction enzymes are a form of __________

A

recombinant DNA technology

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

recombinant DNA technology amplifies, maintains and ________ specific DNA sequences inside the cell

A

manipulates

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

_____________: uses specific enzymes to cut DNA at specific points resulting in double-stranded breaks

A

restriction mapping

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

true/false: if two DNA molecules have sticky ends produced by the same restriction enzymes, they can be joined together

A

true

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

_________: stick to complementary base-pair sequence

A

sticky ends

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

what are the main steps of restriction mapping?

A
  1. digest DNA with restriction enzymes (cut)
  2. separate fragments with electrophoresis
  3. visualize DNA
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
18
Q

_______: is a carrier fragment of DNA

A

vector

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

molecular cloning is a technique that inserts restriction fragments into a vector, why do we do this?

A

when we want to study what a gene actually DOES

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

a vector is inserted unto an organism (usually bacteria), which then amplifies the _____________ within the cell, making _________

A

recombinant DNA molecule

DNA clones

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

what kind of forensic analysis can be done through genetic techniques?

A

DNA fingerprinting/DNA profiling

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

what does DNA fingerprinting/DNA profiling use to match people to DNA?

A

short tandem repeats (STR)

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

what are the four criteria of the combined DNA index system?

A
  1. must have a known chromosome location that independently assorts from all other CODIS STR’s
  2. must have multiple alleles per STR locus in all populations examined
  3. STR alleles must be consistently, reliably, and accurately able to be amplified via PCR
  4. PCR products must distinguish alleles from each other clearly enough for the automated PCR amplification and gel electrophoresis to reliably ID each allele
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
24
Q

in paternity testing, a _______ is calculated for each gene tested (x/y, the probability that a trait actually came from the father in question)

A

paternity index (PI)

25
Q

all paternity index’s are added up to be a __________, which represents all genes analyzed

A

combined paternity index (CPI)

26
Q

________: changing the nucleotide sequence at a specific chromosomal locus to any desired sequence

A

gene editing

27
Q

how does gene editing work? what does it use?

A

uses a DNA endonuclease to cleave the DNA at specific target locations within the genome ( results in double-stranded breaks, which allows the genes to be modified)

28
Q

what are the two main DNA repair mechanisms?

A

non-homologous end joining
homology-directed repair

29
Q

________ allows for gene knockouts/knockins

A

non-homologous end joining

30
Q

how does non-homologous end joining work?

A

proteins recognize double-stranded breaks, the protein complex attaches to both broken ends, the ends are trimmed until a blunt end is achieved, DNA ligase glues the blunt ends together

we do lose some nucleotides from the trimming of the DNA

31
Q

_________ allows for gene insertions and/or modifications

A

homology-directed repair

32
Q

how does homology-directed repair work?

A
  1. a double-stranded break occurs
  2. nucleases trim off a portion of the broken strand, Rad51 binds the undamaged chromatid
  3. strand invasion occurs in a displacement loop (unbroken and broken chromatids are woven together)
  4. DNA replication within the D loop synthesizes new DNA using the sister chromatid as a template
  5. DNA strands are trimmed to blunt ends ad are ligated to finish repair
33
Q

true/false: gene editing tools exploit the cell’s natural DNA repair mechanisms

A

true!

34
Q

what are the two ways artificial gene editing can exploit the cells natural repair mechanisms? where do they “attack”?

A

DNA binding domain

incorporated into a complex with an RNA molecule

35
Q

what were the first two gene editing tools called?

A

ZFNs and TALENs

36
Q

true/false: CRISPR/Cas9 was first found in eukaryotes

A

false, it was first found in prokaryotes in the 1990s

37
Q

what does CRISPR stand for?

A

Clustered Regularly Interspersed Short Palindromic Repeats

38
Q

what is the official name for CRISPR-Cas9

A

CRISPR-associated (cas) genes

39
Q

CRISPR/Cas9 is a natural defense mechanism in prokaryotes
1. CRISPR sequences are transcribed into non-coding RNA\
2. The tracer RNA gene is also transcribed into ________ and Cas9 is produced
3. Processed into _______ by cas-encoded Rnasas

A

tracrRNA

crRNAs

40
Q

true/false: tracrRNA and crRNA form a complex with CAS endonucleases to cleave foreign DNA

A

true

41
Q

true/false: the prokaryote can store the cleaved DNA in its genome for future invader recognition or inheritance

A

true, can incorporate it into its CRISPR region

42
Q

true/false: CRISPR can’t be passed down to progeny

A

false! it can!!

43
Q

how have scientists engineered CRISPR to target any sequence of interest? what have they binded together?

A

crRNA and tracrRNA are fused together into single guide RNA (sgRNA), which scientists can then insert their target DNA into, creating a unique crRNA

Cas9 is used as the endonuclease

44
Q

once Cas9 makes its cut, cell DNA repair finishes gene editing, how do the two DNA repair mechanisms do this?

A

homology-directed repair:
uses supplied DNA fragments as the template for DNA synthesis

nonhomologous end joining:
either results in the deletions of one or more nucleotides, but can also introduce insertions of DNA derived from elsewhere in the genome

45
Q

what three domains make up the Cas9 endonuclease?

A

HNH domain, RuvC domain, PAM

46
Q

what does the HNH domain do?

A

cleaves the strand complementary to the crRNA

47
Q

what does the RuvC domain do?

A

cleaves the opposite strand of DNA

48
Q

what does PAM do?

A

protospacer adjacent motif, destabilises the adjacent sequences for crRNA complementary pairing (in Cas9=NGG)

49
Q

what are the three ways to get CRISPR DNA into cells?

A

mediated transformation, infiltration, particle bombardment

50
Q

agrobacterium contains a plasmid (DNA) called ____________ (Ti plasmid)

A

tumor-inducing plasmid, a portion of the plasmid is called transfer DNA (T-DNA), which can transfer to the plant and recombine with the host genome

51
Q

agrobacterium-mediated transformation:

  1. the _____ of the Ti plasmid is transcribed
  2. Virulence proteins are transcribed and translated
  3. Conjugative transfer proteins are transcribed and translated
  4. The T-DNA will insert itself randomly into the nuclear genome
A

T-DNA

52
Q

how is a Ti-plasmid disarmed? to transfer any gene of interest to plants

A

by removing the opine- and tumor-inducing genes from the T-DNA

53
Q

when transferring a gene of interest to a plant, a gene of interest is inserted between the left and right T-DNA birders into a ________________ (a second plasmid)

A

transformation vector

54
Q

true/false: when transferring a gene of interest to a plant, the two plasmids formed are re-inserted back into the agrobacterium they came from

A

true

55
Q

true/false: genes on the disarmed plasmid are transcribed and translated to virulence and conjugation proteins

A

true

56
Q

true/false: the T-DNA is transcribed from the transformation vector and inserted into the plant host nuclear genome

A

true

57
Q

CRISPR/Cas9 is extensively used in plant biology research, most edits are via _________ repair to promote gene knockouts/downs to determine gene function in model plants

A

NHEJ (nonhomologous end joining)

58
Q

_____________(GMO’s) are defined as organisms that have had their DNA changed by artificial means

A

genetically modified organisms

59
Q

why do some countries allow GMO’s? what is the condition that must be met?

A

they are allowed as long as the results COULD’VE been achieved via selective breeding (i.e we could’ve naturally done it with lots of time and effort, no foreign DNA introduced)

Genetics 1 (11 decks)

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