Explanation of sequencing pcr

1. DNA SEQUENCING
PRESENTED BY
SUMAIRA JAWAID

2. Discovery of DNA Structure
• DNA was first isolated by the
Swiss physician Friedrich
Miescher ,1869.
• DNA is a double helix structure
of long polymer made from
repeating units of two
polynucleotide chains called
nucleotides:are adenine (A),
cytosine (C), guanine (G) and
thymine (T) that carrying genetic
information.

3. Introduction:
• DNA sequencing refers to the general laboratory
technique for determining the sequence of
nucleotides.
• The sequence of DNA is key to understanding
the function of genes and other parts of the
genome.
• DNA sequencing is a process of reading.
• There is a method of storing the data (the
printed word), a language to learn, and a process
of understanding the meaning of what has been
written.

4. History of DNA Sequencing :
• 1869:Deoxyribonucleic acid (DNA) was
first discovered and isolated by
Friedrich Miescher.
• 1977:The chain-termination method
developed by Frederick Sanger.
• 1973: Walter Gilbert and Allan Maxam
"DNA sequencing by chemical
degradation.
• 1984:The first full DNA genome to be
sequenced .

5. Principle of DNA sequencing:
• This method is based on the principle that single-
stranded DNA molecules that differ in length by just
a single nucleotide can be separated from one
another using polyacrylamide gel electrophoresis.
• The sequencing to be copied by using chemical
altered bases.

6. Method of DNA Sequencing
Historically their is two method of DNA sequencing.
• The chain-termination method developed by
Frederick Sanger.
• Walter Gilbert and Allan Maxam "DNA sequencing
by chemical degradation.
Modern sequencing equipments use the principle of
sanger Sequencing.

8. General Procedure of Sequencing
• DNA extraction.
• Optimization of primers(60-54 touch down )
• PCR of sample on same program (60-54).
• Run the PCR sample on Gel electrophoresis(1.2%) to
visualize the result of PCR .
• DNA precipitation (short) then check the result.
• Run Sequencing PCR(BigDye).
• Then run Long percipitation.
• Run sample on genetic analyzer for Sequencing .
• Interpretate the result with the help of BLAST,NCBI.....
different Website .

10. Applications of DNA Sequencing
• DNA sequencing reveals the genetic information that is carried in a
particular DNA segment,
• Homologous DNA sequences from different organisms can be compared
for evolutionary analysis between species or populations.
• DNA sequencing can reveal changes in a gene that may cause a disease.
• DNA sequencing has been used in medicine including diagnosis and
treatment of diseases and epidemiology studies.
• Sequencing has the power to revolutionize food safety and sustainable
agriculture including animal, plant and public health, improving
agriculture through effective plant and animal breeding and reducing
the risks from disease outbreaks.
• DNA sequencing can be used for protecting and improving the natural
environment for both humans and wildlife.

11. THANK YOU