Rakshit Project

AISSCE Practical Examination2021-22
Mrs. Rashmi Ojha Rakshit Sukhpal

Sr. Contents
1 Certificate
2 Acknowledgement
3 Introduction
4 Stages involved
5 Eukaryotic DNA
6 Variable number of Tandem Repeats
7 Restriction Fragment Length

Polymorphism
8 Polymerase Chain Reaction
9 Applications of DNA fingerprinting
10 Conclusion
11 Bibliography
This is to clarify that “Rakshit Sukhpal”
student of class 12th science with Roll no.
__________has successfully completed his
chemistry project on “DNA fingerprinting”
under the guidelines of “Mrs. Rashmi Ohja”.
Signature: Date:
Internal Examiner: External Examiner:

I would like to express my special thanks of gratitude
to my teacher ‘Mrs. Rashmi Ojha’ who gave me the
golden opportunity to do this wonderful project of
chemistry. She also helped me in completing my
project which is “DNA fingerprinting”. I came to
know about so many new things.
I am thankful to the principle ‘Fr. Cantius Ligory’ for

valuable guidance and affectionate nature.
I would also like to thank my parents and friends

who helped me a lot in finalizing this project within
the limited time frame and encourage me to do my
work with my full efficiency.
DNA fingerprinting, also called DNA typing,
DNA profiling, genetic fingerprinting,
genotyping, or identity testing, in genetics, method
of isolating and identifying variable elements within
the base-pair sequence of DNA (deoxyribonucleic
acid). It is the process of determining an individual’s
DNA characteristics. DNA analysis intended to
identify a species, rather than an individual, is called
DNA barcoding.
DNA profiling is a forensic technique in criminal
investigations, comparing criminal suspects’ profiles
to DNA evidence so as to assess to likelihood of their
involvement in the crime. It is also used in parentage
testing, to establish immigration eligibility, and in
genealogical and medical research. DNA profiling
has also been used in the study of animal and plant
populations in the fields of zoology, botany, and
agriculture.
• Cells are broken down to release DNA
• DNA strands cut into fragments
• Fragments separated
• Pattern of fragments analysed
1.
A sample collected from the tissue of a living
or dead organism is treated with chemicals and
enzymes to extract the DNA, which is
separated and purified.
Digestion of DNA by restriction

endonucleases. The sections of DNA that are
cut are called restriction fragments. Thisyields
Fragments are separated on the basis of their
size by this process. DNA being negatively
charged moves towards + terminal. Shorter
fragments move faster.
DNA split into strands using alkaline
solution. DNA fragments are then transferred
from gel to filter paper or nylon membrane,
this is called Southern blotting.
Adding radioactive or coloured probe to the
nylon sheet which is complementary to target
sequences. Each probe only sticks to one or
two specific places on the sheet.
Labelled probe DNA should be hybridized

with the complementary sequences located o
nylon membrane for the detection of position
of later. Membrane is washed to remove non
specific binding and clearing of background.
To detect sequences in genome bound with

hybridized radioactive probe on membrane.
Technique involves alignment of hybridised
membrane with X ray film. The X ray alter
development shows multiple number of bands
that looks like bar codes and known as DNA
fingerprints.
THERE ARE THREE MAJOR FRACTIONS OF
EUKARYOTIC DNA
1 SINGLE COPY SEQUENCE: (50-60 % in mammals)
~5% mammalian genome is coding region
remainder is introns, regulatory regions and
spacer DNA.
2 SIMPLE-SEQUENCE: (10-15% in mammals)
“Satellite DNA”
3 MODERATELY REPEATED : (25-40% in mammals)
small percentage of gene families and
tandemly repeated genes mobile elements.
Satellite DNA
Short (< 100 bp) repeats in tandem arrays
Millions of bp in length
Example: centromeric DNA from humans
contains a 171 bp AT-rich repeat called alpha-
satellite DNA.
• Most of the DNA in a chromosome does
not code for a gene.
• These regions contain sequences that
repeat from 20-100 times (ex-
GTCAGTCAGTCAGTCA)
• There are several known variations of the
Short Tandem Repeats (STR).
• Ex- HUMTH01 (repeating AATG) has
seen different variations.
• Scientists can look at 4-6 different STR’s
in the same person.
• The more STR’s identified the higher the
probability of a match.
• VNTR is more commonly used than
RFLP.
Advantages of VNTR
• Less complex patterns in gel
• Can be used on stains that old or have been
exposed to decomposition.
• STR’s are less likely to degrade
➢ Less than 400 bp
➢ In the middle of the chromosome.
Steps in making an RFLP:
1. Add a restriction enzyme to cut the DNA
into fragments – Exact number and size of
fragments produced varies from person to
person.
2. Fragments are separated by
electrophoresis.
3.The smaller fragments travel further than
the large fragments.
4.This creates a DNA fingerprint.
The DNA is amplified by PCR which contains
three vital steps:
• The solution is heated to 95°C to unzip the
double helix DNA structure.
• The solution is cooled to 55°C to allow the
primers to bind to the ends of the DNA.
• The solution is then reheated to 75°C
which is the optimal temperature for the
Taq polymerase to create new copies of
each DNA strand.
• After 30 cycles the DNA is amplified 1

billion times.
1. Helps diagnose disorders in both parental
and new born babies.
2. Forensic science is the use of scientific
knowledge in legal situations.
3. DNA profiles can be used to determine
whether a particular person is the parent of
a child.
4.Identification of carcass of tissues
5.Detection of somatic mutations or cancer
6.Pathogen identification
With a beginning as a mere forensic tool, the
world of DNA fingerprinting has gone a long
way in revealing the genetic identity of living
beings. With massive evolution in concepts
and techniques it has given a lot to various
fields cutting across the spectrum. In plants it
has not only helped in identifying species but
also in defining a new realm in plant
genomics, plant breeding and in conserving
the biodiversity. With world paving way for
development in biotechnology, DNA
fingerprinting promises a very powerful tool
in our future endeavours.
• Slideshare.net
• 1000sciencefairprojects.com
• Biology NCERT class 12

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AISSCE Practical Examination2021-22

Mrs. Rashmi Ojha Rakshit Sukhpal

6 Variable number of Tandem Repeats

7 Restriction Fragment Length

9 Applications of DNA fingerprinting

Internal Examiner: External Examiner:

I am thankful to the principle ‘Fr. Cantius Ligory’ for

I would also like to thank my parents and friends

Digestion of DNA by restriction

Labelled probe DNA should be hybridized

To detect sequences in genome bound with

• After 30 cycles the DNA is amplified 1

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