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Chapter 13 Summary
Chapter 13 Summary
Chapter 13 Summary
13.1. Introduction
- discussed the various techniques commonly used in molecular
biology and genetics.
- study of the behavior of the chromosomes during mitosis and
meiosis.
- development of histological and cytological techniques to study the
movements and structures of chromosomes.
Identification of Chromosomes
a. Based on the position of the centromere and length of the
chromosomes
b. Labeling the chromosomes using fluorescent dyes or radiolabeled
compounds
c. in situ hybridization of specific chromosomes with radiolabels or
fluorescently labeled nucleic acid probes to locate the position of the
specific genes.
13.2.1.
with
autoradiography
for
The regions between the bands are actually the active regions of
the chromatin where more genes are present, but the quantity of DNA
is very low and therefore the histone proteins. That is why they
appeared unstained or colored lightly.
Specialized
Staining
Technique:
FISH
or
Fluorescence
in
situ
hybridization
Banding Patterns
(1) Q banding obtained by treating with fluorochrome or the
fluorescent dye quinacrin; can be identified by a yellow fluorescence of
different intensity
Quinacrin bids those regions which are rich in A-T and G-C, but
fluorescence oly A-T- quinacrin regions; by this method,
heterochroatn regions are labeld 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.
(2) G banding this technique is not a fluorochrome based
pretreatment; well suited for animal cells; resembles the C-banding
technique without pretreatment
A karyotype analysis usually involves blocking cells in nitosis
and staining the condensed chromosomes wth Giemsa dye.
Giemsa Dye stains regions of chromosomes that are rich in the
base pair Adenine (A) and Thymine (T) producing a dark band.
(3) C banding originated from centromeric or constitutive
heterochromatin. The centromere appears as a stained band; this
technique is well suited for the characterization of plant chromosomes.
This technique onvolves an alkaline pretreatment to complete
the depurination of the DNA. The remaining DNA is again
renatured and stained with Giemsa solution consisting of
methylene azure, methylene violet, methylene blue and eosin.
(4) R banding known as reverse banding technique; this technique
results in the staining of genes rich in G C that is typical for
euchromatins.
(5) Hy banding - a common technique used with plant cells; involves
a pretreatment of the cells in which the cells are warmed in the
presence of HCl and stained with acetocarmine.
13.2.3. Karyotyping a valuable research tool used to determine the
chromosome compliment within somatic or cultured cells; used for the
parental diagnosis and detection of variations in the chromosome number
and structure, aberrations, and anomalies, which are the common cause of
many congenital defects and spontaneous abortions.