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INTRODUCTION:-
A chromosome banding pattern is comprised of alternating light and dark stripes, or
bands that appear along its length after being stained with a dye. OR The treatment
of chromosomes to reveal characteristic patterns of horizontal bands like bar codes is
known as chromosomal banding
A unique banding pattern is used to identify each chromosome and to diagnose chromosomal
aberrations, including chromosome breakage, loss, duplication or inverted segments.
1. Q-banding:
• The alternating bands of bright and dull fluorescence were called Q bands. Quinacrine-
bright bands were composed primarily of DNA that was rich in the bases adenine (A) and
thymine (T), and quinacrine-dull bands were composed of DNA that was rich in the bases
guanine (G) and cytosine (C).
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• This banding pattern is obtained by treating with a fluorochrome or the fluorescent dye
quinacrine. They can be identified by a yellow fluorescence of different intensity.
• Most parts of the stained DNA are heterochromatin.
• A-T regions are seen more in heterochromatin than in euchromatin. Therefore, by this
banding method heterochromatin regions are labeled preferentially.
• The characters of the banding regions and the specificity of the fluorochrome are not
exclusively dependent on their affinity to regions rich in A- T, but it depends on the
distribution of A- T and its association with other molecules such as histone proteins.
Advantages:
It is a simple and versatile technique,
It is used where G – band is not acceptable. It is used as a method of identifying
chromosomes in combination with other procedure.
Study of heteromorphism
Study of human Y chromosome
Disadvantages:
Generally associated with any fluorescence technique: impermanence of the
preparations, the tendency to fade during examination.
2. G-banding:
• Giemsa has become the most commonly used stain in cytogenetic analysis. Staining a
metaphase chromosome with a Giemsa stain is referred to as G-banding. This technique
is not a fluorochrome -based pretreatment.
• It is well suited to animal cells. During mitosis, the 23 pairs of human chromosomes
condense and are visible with a light microscope.
• A karyotype analysis usually involves blocking cells in mitosis and staining the
condensed chromosomes with Giemsa dye.
• The dye stains regions of chromosomes that are rich in the base pairs Adenine (A) and
Thymine (T) producing a dark band. The regions of the chromosome that are rich in
guanine and cytosine have little affinity for the dye and remain light.
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• Standard G-band staining techniques allow between 400 and 600 bands to be seen on
metaphase chromosomes.
• Jorge Yunis introduced a technique to synchronize cells so they are held at the same
stage in the cell cycle. Cells are synchronized by making them deficient in folate,
thereby inhibiting DNA synthesis. By rescuing the cells with thymidine, DNA synthesis
is initiated and the timing of the prophase and prometaphase stages of the cell cycle can
be predicted. Yunis's technique allows more bands to be resolved,
as chromosomes produced from either prophase or prometaphase are less condensed
and are thus longer than metaphase chromosomes.
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Applications:
Most widely used principle methods for demonstrating euchromatic bands than
other two.
Chromosome identification,
Chromosome abnormalities; aneuploidy, breakage and rearrangement,
Chromosome of cultured cells,
Chromosome banding and cancer,
Homogeneity of staining regions,
Gene mapping& High resolution banding (microcytogenetics)
Disadvantages:
The ineffectiveness of determining small translocations, detecting
microdeletions & characterizing the chromosomes of cell lines which are
complex.
3. C-banding:
4. R-banding:
• This is known as a reverse banding technique. This technique results in the staining of
areas rich in G-C that is typical for euchromatins.
• R-banding involves pretreating cells with a hot salt solution that denatures DNA that is
rich in adenine and thymine. The chromosomes are then stained with Giemsa.
• G-, Q-, and R-bandings are not observed with plant chromosomes.
5. Hy-banding:
7. DAPI/Distamycin A Staining
9. T-Banding:
• The T bands apparently represent a subset of the R bands because they are smaller that
the corresponding R bands and are more strictly telomeric. (Gustashaw, 1991).