PCR AND ITS TYPE
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PCR (polymerase chain reaction) is a technique used to amplify specific DNA sequences. It involves repeated cycles of heating and cooling of the DNA sample to separate and copy the DNA strands. There are several types of PCR including nested PCR, which increases sensitivity by using two rounds of amplification, and multiplex PCR, which allows simultaneous amplification of multiple DNA sequences. PCR and its variations have widespread applications in medicine, forensics, agriculture, and research by enabling identification of organisms, understanding disease pathology, and selective isolation of DNA fragments.
PCR AND ITS TYPE
- 1. PCR AND ITS TYPES
- 2. CONTENTS ? PCR Definition ? PCR history ? PCR procedure ? PCR types ? PCR applications ? Conclusion ? References
- 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.