[OCTOBER 13, 2021]

KARYOTYPING
Arboleda, Loui; Capule, Jabez; Leyson, Liv; Salvador, Renee; Saulog, Rafael

Medical Biology 34
De La Salle University—Dasmarinas
College of Science and Computer Studies
Biological Sciences Department
Professor: Ms. Kendy Angue

INTRODUCTION

Karyotyping is a process of determining an individual’s chromosomal set, including

the number of chromosomes and any abnormalities, by arranging pictures of
chromosomes. It also refers to a complete set of chromosomes in a species or an
individual organism, as well as a test that identifies or counts them. It processes by taking
digital images of chromosomes, organizing them into a full karyotype, and interpreting the
results in genetic study or diagnosis. The length of chromosomes, the position of the
centromeres, and the location and sizes of the G-bands are all compared through
karyotyping.

METHODOLOGY

Normally in a physical laboratory, a blood sample is retrieved from the patient. The
sample will be added to a growth medium of which will be incubated for a day at 37°C.
Chemicals are added to halt the cells at metaphase, and will then be put to centrifugation,
containing the supernatant to a test tube with added fixative. The sample cells will then
be put to a slide under the microscope and stained. Under the microscope, the scientist
should be able to identify and visualize the chromosomes.
However, in this activity, we are to use an online virtual lab software. The students
are instructed to use karyotyping for three individuals and determine the abnormalities
that could explain the phenotype with the use of an online Karyotyping software to create
a karyotype image of the three individuals.
Initially, the histories of each of the three patients will be analyzed. On each patient,
their chromosomes will be matched to its homologue, arranging each chromosome until
a completed karyogram is achieved. The total chromosome of each patient will be
counted, and the last set of chromosomes will be determined whether the patient is female
(XX) or male (XY). A diagnosis will be drawn should there be an extra chromosome found
on the karyogram.

HUMAN AND MOLECULAR GENETICS LABORATORY | 1

 [OCTOBER 13, 2021]

As an aid for reading chromosomes, we shall use the Genetic Science Learning
Center website to describe whether a chromosome is metacentric, submetacentric, or
acrocentric. Here, we will also determine the definition of trisomy, monosomy and terminal
deletion in predicting genetic disorders as well as examine the different types of
chromosome abnormalities.

DISCUSSION
CHROMOSOME
[1] What causes a dark band on the chromosome; [2] What is a centromere
[1]
Chromosomes are visualized by utilizing Giemsa
staining (G-banding). The Giemsa reagent stains the DNA
and produces chromosomal banding patterns – light-
staining (G-light) and dark-staining (G-dark) regions. Visible
light bands represent regions that are early replicating, which
are rich in guanine and cytosine. Dark bands on the other
hand represent regions that are late replicating. Dark bands
are rich in nucleotides Adenine (A) and Thymine (T).

[2]
A chromosome is composed of two identical structures of
sister chromatid, held together by a centromere. The centromere is
a constricted region that separates the chromosome into p-arm
(short arm) and q-arm (long arm). It also plays a role in providing
a foundation for the kinetochore. The placement of the centromere
is an important consideration to be of visual use to genetic tests.

HUMAN AND MOLECULAR GENETICS LABORATORY | 2

 [OCTOBER 13, 2021]

PATIENT HISTORIES

Three patient case histories were examined, and karyotypes were completed to properly
diagnose missing extra chromosomes.
PATIENT A

HISTORY
INTERPRETATION
The nearly-full-term fetus of a forty-year-old female. Patient A has 47 chromosomes in
Chromosomes were obtained from fetal epithelial total, is a female due to the presence
cells acquired through amniocentesis.
of XX chromosomes and has an
extra chromosome 21. Thus, the
TOTAL CHROMOSOMES 47 interpretation of Patient A’s
karyotype would be 47, XX, +21.
SEX CHROMOSOMES XX
DIAGNOSIS
CHROMOSOME WITH CHROMOSOME According to the findings, Patient A
EXTRA + 21 has Down Syndrome (Trisomy 21)
due to the presence of an extra
DIAGNOSIS TRISOMY 21 chromosome 21.

HUMAN AND MOLECULAR GENETICS LABORATORY | 3

 [OCTOBER 13, 2021]

PATIENT B

HISTORY
INTERPRETATION
Patient B is a 28-year-old male who is trying to Patient B has 47 chromosomes in
identify a cause for his infertility. Chromosomes were total, is a male (XY Chromosome)
obtained from nucleated cells in the patient's blood. with an extra X chromosome, Sex
chromosome has an extra (X)
TOTAL
CHROMOSOMES
47 chromosome Patient B’s Karyotype
is 47, XXY.
SEX
XXY
CHROMOSOMES

CHROMOSOME DIAGNOSIS
SEX CHROMOSOME
WITH EXTRA +
Patient B has Klinefelter's
KLINEFELTER'S Syndrome, one or more extra sex
DIAGNOSIS
SYNDROME chromosomes (i.e., XXY)

HUMAN AND MOLECULAR GENETICS LABORATORY | 4

 [OCTOBER 13, 2021]

PATIENT C

HISTORY
INTERPRETATION
Patient C died shortly after birth, with a multitude of Patient C has 47 chromosomes in
anomalies, including polydactyly and a cleft lip.
total, has XY sex chromosomes, and
Chromosomes were obtained from a tissue sample.
has an extra chromosome 13.
TOTAL Patient C’s karyotype is 47, XY, +13.
47
CHROMOSOMES

SEX
XY DIAGNOSIS
CHROMOSOMES
Patient C has Patau Syndrome
CHROMOSOME (Trisomy 13) due to the presence of
CHROMOSOME 13
WITH EXTRA +
extra chromosome 13.
DIAGNOSIS TRISOMY 13

HUMAN AND MOLECULAR GENETICS LABORATORY | 5

 [OCTOBER 13, 2021]

GENETIC SCIENCE LEARNING CENTER

THREE KEY FEATURES USED TO READ CHROMOSOMES

The three key features to identify chromosomes are their size, banding pattern, and
centromere position. The size is the easiest way to distinguish a chromosome while
banding patterns can be obtained by using Giemsa dye. Furthermore, the centromere
position could be metacentric, submetacentric, acrocentric, and telocentric depending
on the chromosome:
METACENTRIC SUBMETACENTRIC ACROCENTRIC TELOCENTRIC
the centromere is the centromere is the centromere is The centromere is
located at the center located near the located very near at located very close
of the chromosome center of the the end of the to the end of the
so that the arms are chromosome so one chromosome so one chromosome that the
almost equal. arm is a bit shorter arm is much shorter p arms would not, or
than the other. than the other. barely, be visible.

OWN KARYOTYPE
DIFFICULTIES
ENCOUNTERED
The chromosomes look very
much alike which made it
extremely difficult to distinguish.

HUMAN AND MOLECULAR GENETICS LABORATORY | 6

 [OCTOBER 13, 2021]

TRISOMY
It is a condition wherein there is a presence of an extra chromosome, which causes
developmental abnormalities. One example of this is Down Syndrome (Trisomy 21) in
which there is an extra chromosome 21.

MONOSOMY
It is a condition wherein there is an absence of one member of a pair of chromosomes.
One example of this is Turner syndrome wherein a female has only one X chromosome
(X monosomy) rather than two.

TERMINAL DELETION
It is a condition wherein there is a loss of the end of a chromosome. This may
cause some errors in the development of a baby since some of the "instructions" in the
chromosome are missing.

COMMON GENETIC DISORDERS

CRI DU CHAT SYNDROME

Cri Du Chat, also known as “Cry of the cat”, is the genetic disorder that has a deletion
in one of their copies of chromosome 5. The size of its deletion can vary. The
characteristics of a newborn with cri du chat syndrome include a high-pitched cry, a
small head, widely set eyes, and a flattened bridge of the nose. The cry is caused by
the way the larynx (voice box) develops. Babies with the disease are usually small and
have respiratory problems.

HUMAN AND MOLECULAR GENETICS LABORATORY | 7

 [OCTOBER 13, 2021]

TURNER SYNDROME

Turner Syndrome is a genetic disorder that is caused by having a missing or

incomplete X chromosome which only affects females. Turner syndrome affects
growth and sexual development. Without intervention, girls with this disorder are much
shorter than average. And because their ovaries (which produce eggs, as well as the sex
hormones estrogen and progesterone) do not fully develop and mature, most do not
begin puberty on their own.

KLINEFELTER SYNDROME

Klinefelter Syndrome is a genetic condition caused when someone has two X

chromosomes and one Y chromosome. Typically, testes don't fully develop, and the levels
of the hormone testosterone (important for male sexual development) are lower than
average. As adults, nearly all XXY males are unable to make sperm and so cannot have
biological children (Infertility).

HUMAN AND MOLECULAR GENETICS LABORATORY | 8

 [OCTOBER 13, 2021]

WILLIAMS SYNDROME

Williams Syndrome affects a child's growth, physical appearance, cognitive

development, and personality. This happens when genetic material from one of their
copies of chromosome 7 is missing. A patient with this syndrome has reduced
head/brain size, “elfin face” pointy ears, small and short teeth, growth retardation
during infancy, accelerated sexual maturity, depigmentation (in the eye), and
premature graying of the hair. Some behaviors they manifest are hyper sociability,
enhanced gaze attention, decreased aggression, increased attentive behavior.

CONCLUSION

EXTRA CREDIT
jabs

http://bluehawk.monmouth.edu/~bio/karyotypes.htm

HUMAN AND MOLECULAR GENETICS LABORATORY | 9

 [OCTOBER 13, 2021]

REFERENCES (APA FORMAT)

Port T, Cepaitis A. 2017. Ten human genetic traits of simple inheritance. Science Prof
Online: Your Virtual Science Teacher. [accessed 2020 Aug 03]. Available from:
http://www.scienceprofonline.com/genetics/ten-human-genetic-traits-simple-
inheritance.html

Niego, A., & Benítez-Burraco, A. (2019). Williams Syndrome, Human Self-Domestication,

and Language Evolution. Frontiers in Psychology, 10. doi: 10.3389/fpsyg.2019.00521

Karyotyping Activity. (2021). Retrieved October 9, 2021, from Arizona.edu website:

http://www.biology.arizona.edu/human_bio/activities/karyotyping/patient_c/13c-13.html

Centromere. (2021). Retrieved October 9, 2021, from Genome.gov website:

https://www.genome.gov/genetics-glossary/Centromere

Chromosome Segregation: The Role of Centromeres | Learn Science at Scitable. (2013).

Retrieved October 9, 2021, from Nature.com website:
https://www.nature.com/scitable/topicpage/chromosome-segregation-in-mitosis-the-role-
of-242/

Centromere Proteins Research Tools - Creative BioMart - Creative BioMart. (2020).

Retrieved October 9, 2021, from Creativebiomart.net website:
https://www.creativebiomart.net/researcharea-centromere-proteins_921.htm

Sumner, A. T. (2001). Chromosome Banding. Encyclopedia of Genetics, 348–350. doi:

10.1006/rwgn.2001.0204

Anthony JF Griffiths, Miller, J. H., Suzuki, D. T., Lewontin, R. C., & Gelbart, W. M. (2021).
Topography of the chromosome set. Retrieved October 9, 2021, from Nih.gov website:
https://www.ncbi.nlm.nih.gov/books/NBK22050/

HUMAN AND MOLECULAR GENETICS LABORATORY | 10

 [OCTOBER 13, 2021]

HUMAN AND MOLECULAR GENETICS LABORATORY | 11