pcr (polymerase chain reaction) is the technique used to amplify the part of DNA

1. PCR AND ITS TYPES

2. CONTENTS
• PCR Definition
• PCR history
• PCR procedure
• PCR types
• PCR applications
• Conclusion
• References

6. • POLYMERASE CHAIN
REACTION

7. Pcr

8. PCR STEPS

9. PCR STEPS

11. PCR STEPS

12. Cycling conditions for amplifying longer PCR products
Step Time/cycles Temperature
Initial activation step 2 min 95°C
3-step cycling
Denaturation 10 s 94°C
Annealing 1 min 50–68°C*
Extension 1 min/kb
Number of cycles 40 cycles 68°C
End of PCR cycling Indefinite 4°C

14. PCR

16. PCR TYPES

17. PCR TYPE

19. • Overlap-extension PCR or Splicing by overlap
extension (SOEing) :
• Genetic engineering technique
• used to splice together two or more DNA
fragments OR complementary sequences.
• It is the technique enables creation of
specific and long DNA constructs.
• It can also introduce deletions, insertions or
point mutations into a DNA sequence.

20. • Nested PCR
• This PCR increases the sensitivity
• Two sets of primers,
• A double process of amplification .
• The first set of primers allow a first
amplification. The product of this PCR is
subjected to a second PCR using the second
set of primers.
• Primers used in the second PCR are specific
to an internal amplified sequence in the first
PCR. specificity of the first PCR product is
verified with the second one.

22. • Semi quantitative PCR
• An approximation to the relative amount of
nucleic acids present in a sample,
• The markers commonly used are
• Apo A1 and B actin.
• Amplification product is separated by
electrophoresis

23. • Multiplex PCR
• Multiplex PCR is an adaptation of PCR which
allows simultaneous amplification of many
sequences.
• This technique is used for diagnosis of
different diseases
• Multiplex PCR can detect different pathogens
in a single sample.

24. PCR TYPES

25. Applications of PCR and impact on
science
• PCR and its different variations are highlighted
as the most commonly used in laboratories
and research institutes.
• Thus, these have contributed to identification
• characterization of several organisms and
understanding of physiopathology of diverse
• diseases in human, animal and plants.

26. • MEDICINE
• identification of microorganisms
assurance of blood
• Forensic
• As a basic procedure to investigate Deaths
• (paternity testing)
• Evidence from minimal samples of saliva,
semen or other tissue debris

27. AGRICULTURE
• As conventional PCR or qPCR have also
facilitated research in
• Detection of pathogens in plants, animals,
and the environment; understanding of their
• Epidemiology and, development of new
diagnostic tests, treatments or vaccines.

28. • Selective DNA isolation
• Isolation of DNA fragments from genomic DNA
by selective amplification of a specific region of
DNA.
• This use of PCR augments many methods, such as
generating hybridization Probes and DNA cloning
which require larger amounts of DNA,
representing a specific DNA region.
• PCR supplies these techniques with high
amounts of pure DNA, enabling analysis of DNA
samples Possible.