PCR AND ITS TYPE

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

3. POLYMERASE CHAIN REACTION

4. Pcr

5. PCR STEPS

6. PCR STEPS

7. PCR STEPS

8. 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

9. PCR

10. PCR TYPES

11. PCR TYPE

12. 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.

13. 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.

14. 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

15. 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.

16. PCR TYPES

17. 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.

18. 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

19. 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.

20. 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.