Project Report On DNA

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Project Report on
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An exclusive project report on DNA.

This project report will help you to
learn about: 1. Introduction to DNA 2.
Constituents of DNA 3. Molecular
Structure 4. Forms 5. DNA as the
Genetic Material 6. DNA Content 7.
Unique and Repetitive DNA.
Contents:
1. Project Report on Introduction to DNA

2. Project Report on the Constituents of
DNA
3. Project Report on the Molecular
Structure of DNA
4. Project Report on the Forms of DNA
5. Project Report on DNA as the Genetic
Material
6. Project Report on DNA Content
7. Project Report on Unique and Repetitive
DNA
Project Report # 1. Introduction to

DNA: Privacy - Terms
https://www.biologydiscussion.com/dna/project-report-on-dna/38325#:~:text=It consists of a right,base pair is 3.37 Å. 1/49

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The chromosomes are made up of two types

of macromolecules — proteins and nucleic
acids. The nucleic acids are of two types, viz.
deoxyribonucleic acid (DNA) and
ribonucleic acid (RNA). DNA and RNA are
chain-like macro-molecules that function in
the storage and transfer of genetic
information.
They are major components of all cells.

DNA is found predominantly in the nucleus
while RNA is predominant in cytoplasm.
DNA is the genetic material of most
organisms including many viruses. Some
viruses,’ however, have RNA as their genetic
material.
Deoxyribonucleic acid (DNA) is found in

the cells of living organisms except plant
viruses. DNA is double stranded but in
some cases like bacteriophages (e.g., φ x
174) the DNA is single stranded and
remains coiled and enclosed in a protein
coat. DNA may be circular in bacteria,
spirally coiled and un-branched threads in
mitochondria and plastids of eukaryotic
cells.
Project Report # 2. Constituents of

DNA:
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Deoxyribonucleic acid is a long chain poly-

mer (polynucleotide) composed of
monomeric units, called nucleotides. Each
nucleotide is composed of a nucleoside (a
sugar and a base) and a phosphate group.
Chemical analysis of highly purified

DNA have shown that it is made of
four kinds of monomeric building
blocks each of which contains three
types of molecules :
(i) Phosphoric acid
(ii) Sugar molecule, and
(iii) Organic bases.
(i) Phosphoric Acid:
The phosphoric acid (H3PO4) in the nucleic

acid is called phosphate (Fig 4.1).
Phosphoric acid has three reactive hydroxyl
(-OH) groups of which two are involved in
forming sugar phosphate backbone of DNA.
The phosphate makes a nucleotide
negatively charged.
(ii) Sugar Molecule:
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DNA contains a five carbon sugar, hence it

is a pentose sugar (Fig. 4.1). Since one
oxygen atom at the 2′ carbon is missing, Privacy - Terms

hence it gets its name 2′-deoxyribose. Four

of the five carbon atoms plus a single oxy-
gen atom forms a five-membered ring. The
fifth carbon atom is outside the ring and
forms a part of a -CH2 group.
(iii) Organic Bases:
Different types of heterocyclic nitrogen

containing ring compounds are found in the
structure of DNA. They are called simply as
bases because they can combine with H^ in
acidic solution. They are also referred to as
nitrogenous bases due to presence of
nitrogen.
The bases are of two types mainly —

Pyrimidine and Purine.
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(a) Pyrimidine:
Pyrimidine bases are made up of a six-

membered pyrimidine ring which is similar
to the benzene ring except that it contains
nitrogen in place of carbon at the positions
1 and 3. Pyrimidine bases are of 2 types —
thymine and cytosine (Fig. 4.1), commonly
abbreviated as T and C respectively.
(b) Purine:
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Purine is a derivative of pyrimidine. It

consists of a pyrimidine ring and a five-
membered imidazole ring (having nitrogen
at 7 and 9 positions) which are fused
together at 5 and 4 positions. There are two
purine compounds namely – adenine (A)
and guanine (G) (Fig. 4.1).
Rare or minor bases:
In addition to the four common bases (A, T,

G, C), certain other unusual bases of purine
and pyrimidine derivatives, called rare or
minor bases, occur in small amounts in
DNA of some organisms.
In some viruses, uracil occurs in place of

thymine in DNA. The T-even phages contain
5-hydroxy- methyl-cytosine in place of
cytosine. These modifications protect the
viral DNA from degradation by the host cell
endonuclease. Other rare bases in DNA are
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5-methyl-cytosine, N6-methyl- adenine, N2-

methyl-guanine, etc.
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Molar Ratio of Nitrogenous Bases in

DNA (Chargaff Rules, 1955):
(i) The purine and pyrimidine components

occur in equal amounts in a DNA molecule.
(ii) The amounts of adenine (A) is

equivalent to the amount of thymine (T)
and the amount of cytosine is equivalent to
that of guanine (G).
(iii) In DNA, A + G / T + C value is always

one or nearly one.
(iv) The base ratio A +T/G + C may vary in

the DNA of different groups of organisms
but is constant for particular species.
Therefore, this ratio has been used to
identify the DNA from a particular species.
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Project Report # 3. Molecular

Structure of DNA (Watson and Crick’s
Model):
In 1953 Watson and Crick postulated a

three dimensional working model of DNA,
i.e., double helix structure of DNA based on
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the X-ray data of Wilkins and base-

equivalence observed by Chargaff.
Nucleoside:
A base combined with a sugar molecule is

called a nucleoside. When deoxyribose
sugar binds with base, it makes
deoxyribonucleoside. Obviously, in DNA
four different nucleosides are found.
These are:
(i) Deoxycytidine;
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(ii) Deoxythymidine;
(iii) Deoxyadenosine; and
(iv) Deoxyguanosine.
Nucleotide:
A nucleotide is derived from a nucleoside by

addition of a molecule of phosphoric acid.
The phosphate molecule is linked with
sugar molecule at carbon number 5 or at
carbon number 3 (Fig. 4.2).
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The nucleotides in DNA are of 4 types:
(i) Deoxycytidylic acid;
(ii) Deoxythymidylic acid;
(iii) Deoxyadenylic acid;
(iv) Deoxyguanylic acid (Fig. 4.3, Table 4.1).
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Polynucleotide:
A number of deoxyribonucleotides are

covalently linked one by one to form a
polynucleotide chain, i.e.,
deoxyribonucleotide monomer units are
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united through the formation of

phosphodiester bonds (a diester bond is one
which involves two ester bonds). Fig. 4.4.
Double Helix:
Watson and Crick suggested that in a DNA

molecule, there are two such polynucleotide
chains which are coiled about one another
in a spiral.
The two polynucleotide strands are held

together in their helical configuration by
hydrogen bonding between bases in
opposing strands, the resulting base pairs
being stacked between the two chains
perpendicular to the axis of the molecule
like the steps of a spiral stair case (Fig. 4.5).
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The base-pairing is specific; adenine is

always paired with thymine, and guanine is
always paired with cytosine (Fig. 4.6). Thus,
all base-pairs consist of one purine and one
pyrimidine.
The specificity of base-pairing results from

the hydrogen-bonding capacities of the
bases in their normal configurations. In
their most common structural Privacy - Terms

configurations, adenine and thymine form

two hydrogen bonds, guanine and cytosine
form three hydrogen bonds (Fig. 4.7).
The two strands of DNA double helix are

thus said to be complementary (not
identical). This property, that is
complementarity of the two strands, makes
DNA uniquely suited to store and transmit
genetic information. The base-pairs in DNA
are stacked 3.4 Å apart with 10 base-pairs
per turn (360°) of the double-helix.
The sugar- phosphate backbones of the two

complementary strands are antiparallel,
that is, they have opposite chemical
polarity. Privacy - Terms

As one moves unidirectionally along a DNA

double helix, the phosphodiester bonds in
one strand go from a 3′ carbon of one
nucleotide to a 5′ carbon of the adjacent
nucleotide, whereas those in complemen-
tary strand go from a 5′ carbon to a 3′
carbon. This opposite polarity of the
complementary strand is very important in
considering the mechanism of replication of
DNA.
The high degree of stability of DNA double

helices results in part from the large
number of hydrogen bonds between the
base-pairs and in part from the hydrophobic
bonding between the stacked base-pairs.
The planar sides of them are relatively
nonpolar and thus tend to be water
insoluble (hydrophobic).
This hydrophobic core of stacked base-pairs

con- tributes considerable stability to DNA
molecules present in the aqueous
protoplasm’s of living cells.
Project Report # 4. Forms of DNA:
The DNA molecules exhibit a considerable

amount of conformational flexibility. It can
exist in A, B, C, D and Z forms (Table 4.2). B
form (B-DNA) is the structure proposed by
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Watson & Crick and is the native

conformation of DNA in solution.
It consists of a right-handed antiparallel

double helix of sugar-phosphate backbone,
with purine-pyrimidine base-pairs roughly
perpendicular to the axis of the helix. The
tilt of base-pairs of the helix is 6.3°. One
turn of the helix consists of 10 base-pairs.
The rise of the helix per base pair is 3.37 Å.
A form (A-DNA) has 11 base pairs. The base

pairs are considerably tilted from the axis of
the helix. The axial rise is 2.56Å. The helix
observed under conditions of dehydration
and high concentrations of salt is wider and
shorter than B- helix, the distinction
between the major and minor grooves are
reduced.
C form (C-DNA) results by reduction of

hydration of the B form below 66% with
excess of salt still present. The size of helix
of C form DNA is greater than Å type of
DNA but is smaller than B-DNA. It is about
31 Å. There are 9.33 base pairs per turn. The
axial rise of base pairs is 3.32 Å with a
tilting of about 7.8°.
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D form (D-DNA) and E form (E-DNA) are

found rarely as extreme variants. In case of
D- form there are 8 base pairs per turn of
helix. An axial rise of base pairs is 3.03 Å
with tilting of about 16.7°.
In case of E-form, there are 7.5 base pairs

per turn of helix. Z form (Z-DNA) is an
unique left-handed (Fig. 4.8) double helical
form with a zig-zag (Fig. 4.9) sugar-
phosphate backbone in antiparallel
organization. This DNA has been called Z-
DNA.
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Project Report # 5. DNA as the Genetic

Material:
Transformation Experiment:
Transformation experiment was initially

conducted by F. Griffith in 1928 (Fig. 4.21).
The injected a mixture of two strains of
Pneumococcus (Diplococcus pneumoniae)
into mice. One of these two strains, S III
was virulent and other strain R II was non-
virulent (causing no infection).
Heat-killed virulent S III strain when

injected, showed that infectivity after heat
killing is lost. The mice injected with a
mixture of R II (living) and S III (heat
killed) died and virulent Pneumococcus
could be isolated from these mice. This
phenomenon was described as
transformation.
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O. T. Avery, C. M. Macleod and M.

McCarthy repeated Griffith’s experiment in
an in vitro system in order to identify the
transforming principle responsible for
converting non-virulent into virulent type
and reported their results in 1944 (Fig.
4.22). Virulence in Pneumococcus depends
on a polysaccharide capsule which is
present in virulent strain S III and is absent
in non-virulent strain R II.
The cells of non-capsulated type – Rll were

treated with an extract of DNA from
capsulated strain S III. A few cells of S III
type could be isolated from the mixture. Privacy - Terms

This phenomenon of transferring characters

of one strain to another by using a DNA
extract of the former is called
transformation. When the extract was
treated with DNAse (an enzyme which
destroys DNA) this transforming ability was
lost. Proteases (enzymes which destroy pro-
teins) did not affect the transforming
ability. These experiments thus indicated
that DNA and not the protein, is the genetic
material.
Hershey-Chase Experiment:
Additional direct evidence indicating that

DNA is the genetic material was published
in 1952 by A. D. Hershey and M. Chase. The
experiment showed that genetic
information of a particular bacterial virus
(bacteriophage T2) was present in DNA.
Bacteriophage T2 infects the common colon
bacillus E. coli (Fig. 4.23).
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The basis for Hershey-Chase experiment is

that DNA contains phosphorus but no
sulphur, whereas proteins contain sulphur
but no phosphorus.
Thus, Hershey and Chase were able to

specifically label either (1) the phage DNA
by growth in a medium containing
radioactive isotope of phosphorus 32P, in
place of the normal isotope 31P, or (2) phage
protein coats by growth in a medium
containing radioactive sulphur 35S, in place
of the normal isotope 35S (Fig. 4.24).
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When T2 phage particles labelled with 35S

were mixed with E. coli cells for a few
minutes and were then subjected to
shearing forces by placing the infected cells
in a warring blender, it was found that most
of the radioactivity (and thus the proteins)
could be removed from the cells without
affecting progeny phage production.
When T2 phage in which DNA was labelled

with 32P were used, essentially all
radioactivity was found inside the cells, that
is, it was not subjected to removal by
shearing in a blender (Fig. 4.25).
The sheared off phage-coats were separated

from the infected cells by low-speed
centrifugation which pellets (sediments)
cells while leaving phage particles
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suspended. The results indicated that DNA

of the virus enters the host cell, whereas
protein coat remains outside the cell.
Since progeny viruses are produced inside

the cell, Flershy and Chase’s results
indicated that the genetic information
directing the synthesis of both the DNA
molecules and protein coats of the progeny
viruses must be present in the parental
DNA. Moreover, progeny particles were
shown to contain some of the 32P, but none
35S.
Project Report # 6. DNA Content:
C-Value Paradox:
Large variation in DNA content among

species of the same genus as well as among
genera of the same family has been
observed. This wide range of variation in
the nuclear DNA contents among species
within a genus may be partly attributed to
differences in chromosome numbers.
However, species at the same ploidy level

and with the same chromosome number
may either show little or ‘no variation (e.g.,
Hordeum, Avena, etc.) or may exhibit
many-fold differences (e.g., Lathyrus, Vicia,
Helianthus, Crepis, Allium, etc.). This inter
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specific variation may be continuous or

discontinuous.
Intra- specific variation in DNA content has

also been reported in recent years, so that
the variation of DNA among genotypes is a
rule rather than an exception. This variation
has sometimes been used to explain
mechanism of evolution of specific groups.
In general, the huge difference in DNA

amount not necessarily correlated with the
nature and status of the organism, is
principally due to the repetitive DNA
sequences present in higher amount. In the
plant system, nearly 70- 80% of the DNA in
the cereals is repetitive and only a fraction
of the DNA controls the structural genes for
qualitative characters.
In fact, the comparison of the genomes of

different plant species ranging from algae to
angiosperms show marked variation in the
C value.
The haploid un-replicated DNA content of

an individual is described as its C value. C
value is nearly of 600-fold difference within
the angiosperms alone, ranging from 0.2 pg
in the crucifer – Arabidopsis thaliana to
127pg in Fritillaria assavrica, a liliaceous
species. This paradoxical situation in C
value termed as C value paradox is Privacy - Terms

principally attributed to repetitive DNA

content.
In human, of the 3 billion base pairs which

constitute the entire genome, only about
50000 genes are supposed to be present.
More than 40% of the DNA is highly
repetitive. In general, only a fraction of the
total amount of DNA codes for structural
proteins and enzymes, the rest are repeats –
noncoding or coding for non-specific-effect.
Project Report # 7. Unique and

Repetitive DNA:
The chromosomes of prokaryotes contain

DNA molecule with unique (non-repeated)
base-pair sequences, i.e., each gene which is
a linear sequence of few thousand base-
pairs, present only once in the genome. If
prokaryotic chromosomes are broken into
many short fragments, each fragment will
contain a different sequence of base-pairs.
In higher organisms on the other hand, the

unique DNA sequences which are
principally responsible for qualitative
characters are present in much lower
amount than that of the repetitive
sequences. Such unique sequences are
present in the chromosomes of higher
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organism in between repetitive sequences,

controlling enzyme-protein.
The chromosomes of eukaryotes in general

are very complex. Certain base sequences
are repeated many times in the haploid
chromosome complement, sometimes as
many as million times. DNA containing
such repeated sequences, called repetitive
DNA, often representing a major
component of the eukaryotic genome.
Repetitive DNA:
Types of Repetitive DNA:
The discovery of multiple copies of similar

DNA sequences in chromosomes noted by
Crick is a major event in the study of chro-
mosome research. These repeats may be
highly homogeneous, as in satellite DNA
sequences in Xenopus, or may be moderate
or minor in nature.
These sequences may also be inverted as in

palindromes. In the chromosome structure,
the highly homogeneous repeats may be
tandem located in one locus in cluster form,
whereas minor or moderate repeats may be
interspersed located in intercalary positions
or terminal. In tandem repeats, each
sequence arranged adjacent to the other
forming monomeric unit. Privacy - Terms

Tandem repeats are of two types — similar

repeats and complex combinations of
different repeats, interspersed repeats are
highly scattered, i.e., dispersed throughout
the genome along with other sequences.
Dispersed repeats include mobile elements
such as long interspersed nucleotide
element (LINE), short interspersed
nucleotide element (SINE), long terminal
repeats (LTR).
Repeats may be microsatellites (simple

sequence repeats), minisatellites, satellites.
Size of Repetitive DNA:
The length of repetitive sequences may vary

from simple sequence repeats of di-, tri-,
tetra- or hexanucleotides to nucleosome
repeats of 180 bp and up to 10000 bp or
more in rDNA repeats.
Amount of Repetitive DNA:
The demonstration of the repeated

sequences accounts, to a great extent, for
the huge amount of DNA, noted in the
different organisms. In higher organisms
only little amount of the DNA represents
structural genes containing unique, the rest
being amplification of non-coding
sequences or repeats.
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In the biological system, as a whole, the

nuclear DNA content varies amongst the
organisms without any concomitant
increase in the number and structure of
genes.
The importance of repetitive DNA

sequences can be judged from the very fact
that in the human system almost 40% of
DNA is repetitive in nature. In several plant
species, as a rule 72-75% of DNA is
repetitive whereas in Drosophila only 50%
constitutes the sequences.
Location of Repetitive DNA:
Repetitive DNA sequences are present

throughout the chromosomes including in
centromere and telomeres (also may be
pericentromeric and sub-telomeric),
introns, nucleolar organizing regions,
segments of heterochromatin.
In general, in the entire chromosome

structure, normally highly repeated or
homogeneous repeats are located in one
locus, e.g., secondary constriction or
centromere and moderate, minor repeats
are interspersed throughout. Highly
homogeneous repeats have been located in
ribosomal RNA (5S, 18S, 5.8S, 25S) gene
loci and are mostly AT-rich.
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A characteristic repeat (GGGGATT) found

at the telomeres. Accessory (B)
chromosomes which are heterochromatic in
nature, rich in highly repeated sequences.
Large portion of cereal genome is with
interspersion of short repeats. Presence of
repetitive sequence in introns, in
transposons and in flanking regions of
replicon has been noted.
Origin of Repetitive DNA:
The mechanisms suggested for origin of

repeated sequences include salutatory
replication, unequal crossing over,
transposition including insertion. Repeated
sequences are susceptible to change
resulting from amplification, translocation,
deletion and mutation leading to novel
genomic configuration. Sequences in new
environments may undergo patterning and
amplification, leading to new repeat
families.
Functions of Repetitive DNA:
Several non-specific functions involving

cell-nuclear size and volume, chromosome
cycle, generation time, duration of meiosis,
chromatin folding have often been
attributed to repeated sequences.
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Role of interspersed repeats has been sug-

gested for repair synthesis, regulation of
chromosome structure in folding during
pairing, acting as initiation points in
replication, gene expression through
methylation, gene conversion and
compression.
Function attributed to palindromic repeats

at different levels of protein synthesis
includes recognition systems, both at DNA
and RNA levels, involving deletion and
translocation, cleavage sites, termination of
transcription, binding of regulatory proteins
and attachment of chromosomes with each
other for information transfer.
Intron repeats facilitates alternate splicing

or reshuffling of exons and inter-genic
conversion, permit dispersion and promote
genetic diversity through mobility.
Accessory chromosome repeats in plants
are associated with adaptation in different
environmental set up.
Significance of Repetitive DNA:
Some of the simple sequence repeats can

serve as good markers. Certain tandem
repeated sequences may be unique to a
particular species. Their distribution and
copy number help in differentiating various
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linkage groups in the chromosome

complement.
DNA fingerprinting and molecular

hybridization involving repeats have
immense potential in documentation and
analysis of biodiversity, and tracing the
trends in evolution. The sequence, location
and frequency of short tandem repeats
(STR), which is common in plants’ genome,
have role in determining phylogenetic
status of species.
Detection of Repeats Tm and Cot

Value:
For detection of repetitive DNA, the double

stranded DNA is first denatured by heating
into single stranded DNA which is
accompanied with increase in optical
density (hyperchromicity).
The single stranded DNA is then allowed to

cool slowly to cause re-association between
the complementary sequences into double
stranded DNA which accompanies decrease
in optical density (hypochromicity). Tm is
the temperature at which 50% re-
association is achieved.
The formation of double stranded DNA is

actually measured over different values of a
parameter which is described as Cot value Privacy - Terms

(conc. x time). If solution of different DNA

concentrations is to be compared, same Cot
value can be achieved by altering the time
allowed for re-association.
For DNA sample with high proportion of

repetitive DNA, re-association will be faster
and higher degree of re-association
achieved at lower Cot values.
Localization of Repeats:
For localization of repetitive DNA,

chromosome banding and molecular in situ
hybridization (ISH) techniques are being
applied.
Chromosome Banding:
Differential banding patterns of

chromosomes, usually observed at specific
regions on particular levels, were initially
developed for the analysis of human
chromosome segments.
These bands are made visible through low

and high intensity regions under the
fluorescence microscope or as differentially
stained areas under the light microscope.
The methods were then extended first to
different animals and later to plant
chromosomes.
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The protocol for molecular hybridization,

that is, denaturation at the cytological level,
if followed by renaturation and staining
with different dyes, particularly Giemsa,
gives intensely positive reaction at similar
segments of chromosomes which otherwise
show repetitive DNA.
Obviously such treatment is capable of

revealing repetitive segments in
chromosomes. This banding, following
denaturation-renaturation and Giemsa
staining, is termed G-banding.
Earlier, Caspersson and his colleagues had

recorded differential fluorescence of
different chromosome segments following
staining with various fluorochromes
(quinacrine) and observation under the
ultraviolet microscope and had successfully
employed it in formulating a banding
pattern analysis of the human karyotype.
Such bands were referred to as Q-bands.

Other fluorochromes produce banding
patterns as well, e.g., Hoechst 33258 similar
to Q and ethidium bromide the reverse.
These banding patterns are unique for each

chromosome like fingerprints. The bands
are generally consistent for a taxon except
for minor variations. A number of other
chemicals can also produce bands either Privacy - Terms

identical with or different from the

fluorescent ones, based on different
principles.
Such differential banding patterns, usually

observed at specific regions on particular
chromosomes, are being increasingly used
for the identification of chromosomes.
The chromosome band nomenclature,

adopted at the Paris Conference in
1971, recognized the following types of
banding in human chromosomes (Fig.
4.26A):
Q-bands:
By quinacrine staining and fluorescence.
G-bands:
By staining techniques using Giemsa and

related stains after appropriate pre-
treatment.
R-bands or banding reverse to Q-

bands:
By staining with Giemsa after heating to

87°C.
C-bands:
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For constitutive heterochromatin,

demonstrated by the denaturation-re-
association technique.
E-bands:
Produced by enzymic digestion, as classified

by Lejeune (1973), show woolen and
shrunken regions corresponding to the dark
and faint regions of G-banding. The same
nomenclature can be applied to banding
patterns observed in other eukaryotic
chromosomes.
Other banding patterns of

chromosomes include:
CT-bands: Privacy - Terms

The centromeric and telomeric segments

show bands after treatment in barium
hydroxide, incubation and staining in
‘Stains All’ (4, 5, 4′, 5′-dibenzo-3, 3′-diethyl-
9-methyl-thi- acarbocyanine bromide).
N-bands:
At nucleolus-organizing regions, possibly

due to acidic proteins.
O-bands:
With orcein staining, mainly for plant

chromosomes; both intercalary and centro-
meric heterochromatin show bands. All the
above methods of banding bring out clearly
the differentiation of chromosome seg-
ments which can easily be analysed under
the microscope. This technique permits
identification of chromosome segments
with specific molecular complexity such as
repeat sequence.
The comparison of banding pattern between

different genotypes, altered and unaltered,
can localize the segments which have
undergone alterations. The importance of
banding can be judged by the very fact that,
R-banding that is the reverse banding has
been utilized for mapping of gene
sequences in human chromosome.
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In Situ Hybridization (ISH: FISH &

GISH):
Besides the chromosome banding, for

localization and mapping of different
segments of chromosomes and gene loci at
the microscopic level, application of
molecular hybridization in situ is now
widely adopted (Fig. 4.26B).
In situ hybridization (ISH) principally uses

probe sequences, tagged with radioisotopes
or fluorescent compounds (or a chemical
reporter). The initial step is denaturation of
the target which is followed by
hybridization with probe of the
complementary sequences to undergo re-
annealing or pairing.
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The complementary sequences of the probe

bind selectively at the target site. The
hybridized sites are localized either through
autoradiography or immune-fluorescence
as well as counter staining with specific
stains detected cytologically. The in situ
hybridization technique was developed ini-
tially by Pardue and Gall and later modified
by different authors.
The fluorescent in situ hybridization (FISH)

is the most powerful technique at present,
through which the target loci at the
chromosome is hybridized with
complementary probe sequence, tagged
with fluorescent compound. There are two
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approaches, namely direct or indirect, of

FISH technique in plant chromosome.
In the indirect method, the probes are

tagged with reporter molecule, such as
biotin, digoxigenin and finally they are
located by fluorochrome conjugated
antibodies such as avidin.
The main principle of this method is to

make the probe- target sequence as
antigenic so that, it can be detected through
antibody. The common fluorochromes are
FITC (fluorescein-isothiocyanate) as well as
rhodamine. In the direct method, the probe
is directly labelled by fluorochrome-
labelled antibodies. The direct labelling of
fluorochrome with the probe is the rapid
method of ensuring good resolution.
Due to lack of karyomorphological markers,

metaphase chromosome analysis cannot
distinguish parental genomes in hybrids. If
ISH technique is applied with total genomic
probes where the plant has multi-genomic
constitution, the parental chromosome can
be directly identified in the hybrids, such as
Triticum and Secale in Triticale.
Thus method of total genome in situ

hybridization is otherwise termed as GISH
(genomic in situ hybridization) technique.
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Since its first demonstration in

identification of parental genomes in hybrid
between Hordeum chilense and Secale
africanum by Schwarzacher et al., the
technique has been extensively applied to
elucidate ancestry of hybrids and
polyploids. GISH remains a very effective
tool in genome identification, their
orientation and in establishing genomic
relationships between species.
The use of GISH in meiosis helps in

understanding inter-genomic homologies as
well as in elucidating the possible transfer
of chromosome segments through inter-
genomic recombination.
The FISH technique, using different colour

combinations by different probes, is now
being applied to detect simultaneously
different genomes, or chromosome
segments by extension of the technique –
otherwise termed as multi-colour FISH.
This method has been used to distinguish
three genomes in hexaploid wheat and to
detect several translocation sites and
insertions in polyploid species of Triticum
and Aegilops.
The multi-colour FISH technique is now

regarded as a powerful tool for gene
mapping as well as detection of
abnormalities, including insertion and Privacy - Terms

breakage points with chromosome specific

or genome specific dispersed probes. The
term chromosome painting is used in FISH
technique where chromosome specific
dispersed probes are used to detect the
location of complementary target sequences
in the complement.
The FISH technique in recent years has

further been modified to locate single copy
or tandem sequences, utilizing primer
mediated extension and amplification in
situ by PGR method. The application of this
method lies also in confirmation of location
of foreign gene at the chromosome level in
transgenic individuals.
Related Articles:
1. Science Fair Project on DNA

Fingerprinting
2. Top 5 Techniques of Chromosome
Banding
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3/1/22, 2:44 PM Project Report on DNA

An exclusive project report on DNA.

1. Project Report on Introduction to DNA

Project Report # 1. Introduction to

https://www.biologydiscussion.com/dna/project-report-on-dna/38325#:~:text=It consists of a right,base pair is 3.37 Å. 1/49

The chromosomes are made up of two types

They are major com­ponents of all cells.

Deoxyribonucleic acid (DNA) is found in

Project Report # 2. Constituents of

https://www.biologydiscussion.com/dna/project-report-on-dna/38325#:~:text=It consists of a right,base pair is 3.37 Å. 2/49

Deoxyribonucleic acid is a long chain poly­-

Chemical analysis of highly purified

(i) Phosphoric acid

(ii) Sugar molecule, and

(iii) Organic bases.

(i) Phosphoric Acid:

The phosphoric acid (H3PO4) in the nucleic

(ii) Sugar Molecule:

DNA contains a five carbon sugar, hence it

https://www.biologydiscussion.com/dna/project-report-on-dna/38325#:~:text=It consists of a right,base pair is 3.37 Å. 3/49

hence it gets its name 2′-deoxyribose. Four

(iii) Organic Bases:

Different types of hetero­cyclic nitrogen

The bases are of two types mainly —

Pyrimidine bases are made up of a six-

https://www.biologydiscussion.com/dna/project-report-on-dna/38325#:~:text=It consists of a right,base pair is 3.37 Å. 4/49

Purine is a derivative of pyrimidine. It

Rare or minor bases:

In addition to the four common bases (A, T,

In some viruses, uracil occurs in place of

https://www.biologydiscussion.com/dna/project-report-on-dna/38325#:~:text=It consists of a right,base pair is 3.37 Å. 5/49

5-methyl-cytosine, N6-methyl- adenine, N2-

Molar Ratio of Nitrogenous Bases in

(i) The purine and pyrimidine components

(ii) The amounts of adenine (A) is

(iii) In DNA, A + G / T + C value is always

(iv) The base ratio A +T/G + C may vary in

Project Report # 3. Molecular

In 1953 Watson and Crick postulated a

https://www.biologydiscussion.com/dna/project-report-on-dna/38325#:~:text=It consists of a right,base pair is 3.37 Å. 6/49

the X-ray data of Wilkins and base-

A base combined with a sugar molecule is

(iii) Deoxyadenosine; and

A nucleotide is derived from a nucleoside by

https://www.biologydiscussion.com/dna/project-report-on-dna/38325#:~:text=It consists of a right,base pair is 3.37 Å. 7/49

The nucleotides in DNA are of 4 types:

(i) Deoxycytidylic acid;

(ii) Deoxythymidylic acid;

(iii) Deoxyadenylic acid;

(iv) Deoxyguanylic acid (Fig. 4.3, Table 4.1).

https://www.biologydiscussion.com/dna/project-report-on-dna/38325#:~:text=It consists of a right,base pair is 3.37 Å. 8/49

A number of deoxyribonucleotides are

https://www.biologydiscussion.com/dna/project-report-on-dna/38325#:~:text=It consists of a right,base pair is 3.37 Å. 9/49

united through the formation of

Watson and Crick suggested that in a DNA

The two polynucleotide strands are held

https://www.biologydiscussion.com/dna/project-report-on-dna/38325#:~:text=It consists of a right,base pair is 3.37 Å. 10/49

The base-pairing is specific; adenine is

The specificity of base-pairing results from

https://www.biologydiscussion.com/dna/project-report-on-dna/38325#:~:text=It consists of a right,base pair is 3.37 Å. 11/49

configurations, adenine and thymine form

The two strands of DNA double helix are

The sugar- phosphate backbones of the two

They are major components of all cells.

Deoxyribonucleic acid is a long chain poly-

Different types of heterocyclic nitrogen

In fact, the comparison of the genomes of

principally attributed to repetitive DNA

The chromosomes of prokaryotes contain

In the biological system, as a whole, the