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MT 6315 > UNIT 7: KARYOTYPING > Flashcards

UNIT 7: KARYOTYPING Flashcards

1
Q

is the number and appearance of chromosome in the nucleus of a eukaryotic cell

A

Karyotype

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2
Q

Information on the karyotype

A

SiSi Po Pre

Size of chromosome
Size of satellites
Position of centromere
Presence of secondary constrictions

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3
Q

T or F: Karyotyping begins with the culturing of cells

A

T

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4
Q

Karyotype comes from the Greek word
_______ which means ________.

A

Karyon; nucleus

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5
Q

the study of whole sets of chromosomes

A

Karyology

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6
Q

the standard
format of representing chromosomes as
diagram when the haploid set of
chromosomes of an organism are ordered
in a series of decreasing size

A

Idiogram or Karyogram

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7
Q

show larger differences between smaller and larger chromosomes in a set

have more acrocentric chromosomes and
relatively advanced feature

A

Asymmetric karyotype

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8
Q

show lesser difference between smaller and larger chromosomes in a set

have more metaphase chromosomes and no advanced feature.

A

Symmetric karyotype

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9
Q

In 1931 ______________, a Russian scientist suggested that in flowering plants there is a predominant trend towards karyotype asymmetry.

This trend has been carefully studied in
the genus _______ of the family compositae.

A

G.A. Levitzky; Crepis

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10
Q

T or F: Species showing a greater asymmetry is less advanced.

A

F

It is more advanced.

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11
Q

Proportion of metacentric, acrocentric chromosomes in a set.

Ratio between size of largest and smallest chromosomes in a set.

A

Degree of asymmetry

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12
Q

The higher the proportion of __________ chromosomes, the greater the value
of size ratio, more asymmetrical is a karyotype

A

acrocentric

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13
Q

is the process of pairing and ordering all the chromosomes
of an organism, thus providing a genome-wide snapshot of an
individual’s chromosomes.

A

Karyotyping

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14
Q

T or F: Karyotypes are prepared using standardized staining procedures that
reveal characteristic structural features for each protein.

A

F

They reveal features of each chromosome

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15
Q

Banding patterns are a product of what procedure?

A

staining

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16
Q

T or F: Karyotyping analysis focuses more on aneuploidies rather than aberrations

A

T

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17
Q

T or F: Telomeres can also be present in ring chromosomes

A

F

Their presence serves as a clue that you are looking at a linear chromosome

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18
Q

Euchromatin or Heterochromatin?

More proteins

A

Heterochromatin

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19
Q

Euchromatin or Heterochromatin?

Transcriptionally active

A

Euchromatin

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20
Q

Euchromatin or Heterochromatin?

Light staining

A

Euchromatin

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21
Q

Euchromatin or Heterochromatin?

Abundant GC bonds

A

Euchromatin

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22
Q

Euchromatin or Heterochromatin?

Abundant AT bonds

A

Heterochromatin

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23
Q

Causes nucleosomes to pack tightly together

A

Methylation of DNA and histones

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24
Q

T or F: In methylation, transcription factors cannot bind the DNA, so genes are not expressed.

A

T

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25
Q

Results in loose packing of nucleosomes

A

Acetylation of histones

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26
Q

T or F: In acetylation, transcription factors can bind the DNA, so genes are expressed.

A

T

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27
Q

Materials needed for karyotyping

A

Sterile 5mL syringe
21G syringe needle
15mL conical tubes
Green-top vacutube
Glass slides
Pasteur Pipette
Pipettor and Pipette tips
Serological Pipettes

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28
Q

Reagents needed for karyotyping

A

Glacial acetic acid
Methanol
KCl (hypotonic solution)
RPMI Growth Medium
Fetal Bovine Serum
Phytohemagglutinin
Colcemid
Giemsa Dye
Trypsin

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29
Q

Equipment needed for karyotyping

A

Centrifuge
Incubator at 37°C CO2
Refrigerator
Inverted Microscope

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30
Q

5 Major steps in karyotyping

A
  1. Short term lymphocyte culture
  2. Harvesting of lymphocytes
  3. Fixing the cells
  4. Making the chromosome slides
  5. Slide analysis
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31
Q

T or F: The collected blood will be grown in vivo by adding cell culture growth medium, fetal bovine serum, antibiotics, and phytohemagglutinin (PHA) – the reagent that induces mitotic activity

A

F

It is grown in vitro

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32
Q

T or F: Hypotonic medium is used for lysing the cells

A

T

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33
Q

What reagent ensures that only lymphocytes will be affected by the culture process?

A

RPMI

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34
Q

Mitogen; Reagent that stimulates mitosis

A

Phytohemagglutinin

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35
Q

Arrests mitosis at metaphase

A

Colcemid/Colchicine

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36
Q

Reagent used to digest proteins

A

Trypsin

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37
Q

T or F: In step 1, the cultured blood cells will be grown at 37 °C incubator for 3 days

A

T

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38
Q

In step 1, the cells must be in ____________ phase because splitting of a cell line 2 days before harvesting, and changing the medium 1 day before harvesting, stimulates cell proliferation significantly.

A

logarithmic

39
Q

T or F: In step 2, addition of pre-cooled colcemid arrests the cell cycle at metaphase stage

A

F

It should be pre-warmed

40
Q

Incubation with colcemid should be done for ____mins

A

15

41
Q

Optimal exposure time to colcemid requires a balance between

A

proliferative activity index of cells and concentration of colcemid

42
Q

Short proliferative index requires a ______ _______ ________ of colcemid

A

short high burst (flash)

43
Q

This refers to how fast or slow the cells grow

A

proliferative activity index of cells

44
Q

In step 2, _____ ________ will be formed by centrifuging the tube at 1000 RPM for 10 mins

A

cell pellets

45
Q

T or F: Using Actinomycin D or ethidium
bromide (added before harvesting), or bromodeoxyuridine (BrdU) before colcemid treatment is just optional

A

T

46
Q

can significantly increase the total yield of
metaphase chromosomes

A

Cell synchronization

47
Q

Cells are arrested at ___ phase by adding an excess amount of BrdU overnight (16 h).

A

S

48
Q

In step 2, what is added after arresting the cells at S phase?

A

Thymidine (for 5.5 h before colcemid treatment)

49
Q

T or F: In step 3, the cell suspension in hypotonic state will be centrifuged for 1200 RPM for 10 mins.

A

F

Only for 5 mins

50
Q

ratio of absolute methanol to glacial acetic acid

A

3:1

51
Q

the most common method of staining chromosomes for differentiation

A

GTG-banding

52
Q

Meaning of GTG in GTG-banding

A

G-bands by Trypsin using Giemsa

53
Q

GTG-banding uses trypsin that digests the chromosomes at regions rich in basic amino acids, specifically ____ and ____

A

Arg; Lys

54
Q

In step 5, slides that will be chosen for analysis and visualization must be:

A

Properly trypsinized chromosomes
Clearly defined metaphase spreading

55
Q

is a part of a chromosome which is
clearly distinguishable from its adjacent
segments by appearing darker or lighter
with various banding methods (Paris
conference, 1971)

A

Band

56
Q

T or F: Classical cytogenetics provide low resolution

A

T

57
Q

T or F: Changing the stain would not affect the banding pattern

A

F

Changing the stain would also change the banding pattern

58
Q

In 1958, _________ et al., published there first paper describing the use of quinacrine mustard to stain chromosome there by ushered in a new era of chromosome banding

A

Caspersson

59
Q

the first attempt to provide
nomenclature for chromosome banding in any species and thus its
recommendations have been adopted to nonhuman species as well

A

Paris report (1971)

60
Q

T or F: Studying banding patterns allows you to see smaller pieces of the chromosome, so that you could identify smaller structural chromosome abnormalities not visible on a routine analysis.

A

T

61
Q

Classification of Banding Techniques is based on

A

GC and AT rich regions
Constitutive Heterochromatin Region

62
Q

Classification of Banding Techniques

A

Quinacrine 1958 by Casperson et al.
Giemsa 1971 by Summer et al.
NOR 1973 by Matsui & Sasaki
Centromeric 1978 by Linde & Laursen

63
Q

Giemsa stain reacts with

A

Sulfate structures

64
Q

T or F: Sulfate structures are found in sugar and phosphate groups of DNA

A

F

They are found in proteins

65
Q

T or F: Trypsinization precedes staining

A

T

66
Q

T or F: Darkness and lightness of Giemsa stain have nothing to do with the number of Hydrogen bonds between bases

A

T

67
Q

Banding based on fluorescence

A

Q-banding

68
Q

In G-banding, ___-rich regions stain dark due to abundance of proteins

A

AT

69
Q

Quinacrine fluotescent dye stains ___-rich regions

A

AT

70
Q

Banding pattern is opposite to G-banding

A

R-Banding

71
Q

Type of banding that uses phosphate buffer

A

R-banding

72
Q

C in C-banding stands for

A

Centromeric OR Constitutive

73
Q

Usually stains the entire long arm of the Y chromosome

A

C-banding

74
Q

T or F: Human chromosomes do not need a lot of protein encoding regimens

A

T

75
Q

Type of bending wherein euchromatin stains dark and heterochromatin stains light

A

R-banding

76
Q

Type of banding done by denaturation with alkaline (strong base) followed by Giemsa

A

C-banding

77
Q

Type of banding that involves silver staining regions

A

NOR banding

78
Q

NOR stands for

A

Nucleolar Organizing Regions

79
Q

First stain used in Q banding

A

Quinacrine Mustard

80
Q

In Q banding, stained chromosomes need to be subjected to ____ ________ for excitation

A

UV light

81
Q

In Q banding, ____ regions quench dye out and do not fluorescence

A

GC

82
Q

Type of banding used in study of chromosome heteromorphism

A

Q banding

83
Q

T of F: Q banding does not fade during examination

A

F

Its disadvantage is its tendency to fade

84
Q

In G banding, interaction of the DNA with _______ & _______ components of stain brightens sulfur rich regions

A

thiazine; eosin

85
Q

Type of banding used in identification of bands rich in sulfur content

A

G banding

86
Q

Type of banding used in the identification of chromosomal abnormalities and gene mapping

A

G banding

87
Q

T or F: G banding is also used in plants

A

F

It is not used in plants

88
Q

T or F: In C banding, repetitive DNA renature but unique DNA do not renature

A

T

89
Q

T or F: C banding is used in paternity testing and gene mapping

A

T

90
Q

Each chromosome arm is divided into regions, or _________ ________, that can be seen using a microscope and special stains.

A

cytogenetic bands

91
Q

T or F: The cytogenetic bands are labeled p1, p2, q1, q2, etc., counting from the centromere out toward the telomeres.

A

T

92
Q

T or F: Following ISCN, each area of chromosome has a given number.

A

T

93
Q

Each human chromosome has a short arm and long arm, separated by a ___________

A

centromere

MT 6315 (2 decks)

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