Karyotype

The study of structure and

function of chromosomes.
DO NOT CUT OUT ALL CHROMOSOMES!!! or you will lose chromosomes.

Cut out only one at a time AND THEN ATTEMPT TO MATCH THEM
Chromosome banding technique:
Types of chromosome banding technique:
Q-banding:
● Q banding used quinacrine stain (quinacrine dihydrochloride or quinacrine mustard) and it is the simplest and
the first chromosomal banding method. Quinacrine stained chromosomes show a characteristic pattern of
bright bands on a darker background. Because quinacrine is a DNA intercalating agent and a fluorescent
compound, the bands appear only when the chromosomes are exposed to ultraviolet (UV) light.
● Ultraviolet light causes the quinacrine molecules to shine, therefore parts of the chromosome intercalated
with quinacrine shine brightly, whereas other parts remain dark.
● This bright-dark banding pattern is highly reproducible and is also specific for each chromosome. Thus with
quinacrine banding is used to identify particular chromosomes in a cell, and also to determine if a
chromosome is structurally abnormal.
● Quinacrine is an intercalating agent and it intercalate between the base pairs of the DNA helix. Quinacrine
has more affinity for DNA sequence containing AT sequence. Therefore fluorescence of quinacrine is
enhanced along AT rich sequence and appear bright than GC rich sequence.
● * quinacrine is carcinogenic.
Types of chromosome banding technique:

G-banding:
● G-banding is the most frequently used technique for chromosome staining in cytogenetics.
● Giemsa staining is an excellent non fluorescent staining techniques.
● Giemsa also creates a reproducible pattern of bands on each chromosome. It is still not clear why
chromosomes show bands when they are stained with quinacrine or Giemsa
● Bright field microscope is used for visualization
● In G banding technique, before using Giemsa stain, there is always a pretreatment step.
● Usually proteolytic enzyme trypsin is used for pretreatment. Therefore the process is also known as GTG
banding (G-banding by trypsin with Giemsa).
● There is an alternative to Giemsa stain and it is Wright stain.
● G-banding also produce same banding pattern as Q banding along the length of chromosome. Geimsa stain
has more affinity for DNA sequence rich in AT content hence stained dark while sequence rich in GC content
stain light.
G-banding:
Types of chromosome banding technique:

C-banding:
● C-banding is called Centromeric heterochromatin staining
● In this technique, before using Giemsa stain the cell is pretreated with alkali. Therefore this technique is also
known as CBG-staining (B-banding by base with Giemsa).
● Centromeric heterochromatin and distal part of Y chromosome containing highly repetitive DNA sequence
(satellite DNA) is used to stain by C-banding technique.
● After staining, banding is viewed by bright-field microscope.
C-banding:
Types of chromosome banding technique:

R-banding:
● R-banding is called Reverse chromosome banding.
● With this banding technique the band pattern produced in chromosome is reversed to the band produced by
G-banding and Q-banding. ie. The dark band (AT rich region) observed in G-banding technique appears light in
R-banding and vice versa.
● R-banding technique also uses Giemsa stain but before staining with Giemsa the slide is heated at 88°C in a buffer
solution. Heating causes denaturation of DNA.
● Denaturation of chromosome at AT rich region occurs at faster rate resulting in loss of DNA from these regions but
not from GC rich region. The GC rich region is then stained by Giemsa stain which appears stained (R band)
● G-banding is usually preferred over R-banding. However R-banding can be used for chromosome identification.
● In some cases, R-banding is a useful complement to G-banding because some small light G band can be more
easily detected when they are stained by R-banding.
● R-banding is also useful for visualization of telomere sequence at the ends of chromosomes. Telomeres stained
dark with R-banding while light with G-banding.
Chromosome banding technique:
What is a Karyotype?
Karyotype
● Karyotype is the actual picture of total chromosomes
of a cell's nucleus.
● It consists of a whole set of homologous
chromosome pairs, arranged in decreasing series of
their size.
● Microphotographs of metaphase chromosomes help
in lining up of the homologous pairs and the
preparation of a karyotype.
● By analyzing the karyotype of the organism, it is
possible to detect genetic disorders and other
information about the individual. It also tells the
gender of the individual.
 Idiogram
● Idiogram is a schematic diagram of a karyotype that illustrates all chromosome maps. It is not an actual picture of
total chromosomes of a cell.
● Idiogram shows the chromosome maps indicating the locations of genes as bands,provides locations of individual
genes present in a chromosome.
● Idiograms are useful in identifying various abnormalities associated with a range of chromosomal disorders and
determining the links between structural abnormalities and individual genes correlated with various diseases or
syndromes.