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Pcr pcr applications

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RAHUL GAUTAM

Polymerase chain reaction (PCR) amplifies a specific DNA sequence in vitro. It involves repeated cycles of heating and cooling DNA in the presence of primers, DNA polymerase, and nucleotides. Each cycle doubles the number of DNA copies. After many cycles, a single copy of DNA can be amplified exponentially to generate millions of copies. PCR has numerous applications including disease diagnosis, genetic fingerprinting, gene cloning, forensics, and molecular biology research.

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Polymerase Chain ReactionPolymerase Chain Reaction (PCR) and Its Applications(PCR) and Its Applications by RAHUL GAUTAM
What is PCR?What is PCR? It was invented in 1983 by Dr. Kary Mullis, for which he received the Nobel Prize in Chemistry in 1993. PCR is an exponentially progressing synthesis of the defined target DNA sequences in vitro.
What is PCR? :What is PCR? : Why ¡°Polymerase¡±?Why ¡°Polymerase¡±? It is called ¡°polymerase¡± because the only enzyme used in this reaction is DNA polymerase.
What is PCR? :What is PCR? : Why ¡°Chain¡±?Why ¡°Chain¡±? It is called ¡°chain¡± because the products of the first reaction become substrates of the following one, and so on.
Pcr pcr applications
What is PCR? :What is PCR? : The ¡°Reaction¡± ComponentsThe ¡°Reaction¡± Components 1) Target DNA - contains the sequence to be amplified. 2) Pair of Primers - oligonucleotides that define the sequence to be amplified. 3) dNTPs - deoxynucleotidetriphosphates: DNA building blocks. 4) Thermostable DNA Polymerase - enzyme that catalyzes the reaction 5) Mg++ ions - cofactor of the enzyme 6) Buffer solution ¨C maintains pH and ionic strength of the reaction solution suitable for the activity of the enzyme
The ReactionThe Reaction THERMOCYCLERPCR tube
Pcr pcr applications
Pcr pcr applications
Pcr pcr applications
DNA copies vs Cycle number 0 500000 1000000 1500000 2000000 2500000 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 Cycle number DNAcopies
Pcr pcr applications
Pcr pcr applications
Applications of PCRApplications of PCR ? Classification of organisms ? Genotyping ? Molecular archaeology ? Mutagenesis ? Mutation detection ? Sequencing ? Cancer research ? Detection of pathogens ? DNA fingerprinting ? Drug discovery ? Genetic matching ? Genetic engineering ? Pre-natal diagnosis
Applications of PCR Basic Research Applied Research ? Genetic matching ? Detection of pathogens ? Pre-natal diagnosis ? DNA fingerprinting ? Gene therapy ? Mutation screening ? Drug discovery ? Classification of organisms ? Genotyping ? Molecular Archaeology ? Molecular Epidemiology ? Molecular Ecology ? Bioinformatics ? Genomic cloning ? Site-directed mutagenesis ? Gene expression studies
Applications of PCR Molecular Identification Sequencing Genetic Engineering ? Molecular Archaeology ? Molecular Epidemiology ? Molecular Ecology ? DNA fingerprinting ? Classification of organisms ? Genotyping ? Pre-natal diagnosis ? Mutation screening ? Drug discovery ? Genetic matching ? Detection of pathogens ? Bioinformatics ? Genomic cloning ? Human Genome Project ? Site-directed mutagenesis ? Gene expression studies
APPLICATIONS OF REAL-TIME PCR
MMOLECULAROLECULAR IIDENTIFICATION:DENTIFICATION:
Pcr pcr applications
Detection of Unknown MutationsDetection of Unknown Mutations Molecular Identification:
Classification of OrganismsClassification of Organisms 1) Relating to each other 2) Similarities 3) Differences * Fossils * Trace amounts * Small organisms ! DNA ! Molecular Identification: Insufficient data
Pcr pcr applications
Pcr pcr applications
Detection Of PathogensDetection Of Pathogens Molecular Identification:
Pcr pcr applications
SSEQUENCINGEQUENCING Nucleotides (dNTP) are modified (dideoxynucleotides = ddNTP) NO polymerisation after a dideoxynucleotide! Fragments of DNA differing only by one nucleotide are generated Nucleotides are either or
Pcr pcr applications
SummarySummary blood, chorionic villus, amniotic fluid, semen, hair root, saliva 68,719,476,736 copies Gel Analysis, Restriction Digestion, Sequencing
ConclusionConclusion The speedspeed and easeease of use, sensitivitysensitivity, specificityspecificity and robustnessrobustness of PCR has revolutionised molecular biology and made PCR the most widely used and powerful technique with great spectrum of research and diagnostic applications.

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Pcr pcr applications

  • 1. Polymerase Chain ReactionPolymerase Chain Reaction (PCR) and Its Applications(PCR) and Its Applications by RAHUL GAUTAM
  • 2. What is PCR?What is PCR? It was invented in 1983 by Dr. Kary Mullis, for which he received the Nobel Prize in Chemistry in 1993. PCR is an exponentially progressing synthesis of the defined target DNA sequences in vitro.
  • 3. What is PCR? :What is PCR? : Why ¡°Polymerase¡±?Why ¡°Polymerase¡±? It is called ¡°polymerase¡± because the only enzyme used in this reaction is DNA polymerase.
  • 4. What is PCR? :What is PCR? : Why ¡°Chain¡±?Why ¡°Chain¡±? It is called ¡°chain¡± because the products of the first reaction become substrates of the following one, and so on.
  • 6. What is PCR? :What is PCR? : The ¡°Reaction¡± ComponentsThe ¡°Reaction¡± Components 1) Target DNA - contains the sequence to be amplified. 2) Pair of Primers - oligonucleotides that define the sequence to be amplified. 3) dNTPs - deoxynucleotidetriphosphates: DNA building blocks. 4) Thermostable DNA Polymerase - enzyme that catalyzes the reaction 5) Mg++ ions - cofactor of the enzyme 6) Buffer solution ¨C maintains pH and ionic strength of the reaction solution suitable for the activity of the enzyme
  • 11. DNA copies vs Cycle number 0 500000 1000000 1500000 2000000 2500000 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 Cycle number DNAcopies
  • 14. Applications of PCRApplications of PCR ? Classification of organisms ? Genotyping ? Molecular archaeology ? Mutagenesis ? Mutation detection ? Sequencing ? Cancer research ? Detection of pathogens ? DNA fingerprinting ? Drug discovery ? Genetic matching ? Genetic engineering ? Pre-natal diagnosis
  • 15. Applications of PCR Basic Research Applied Research ? Genetic matching ? Detection of pathogens ? Pre-natal diagnosis ? DNA fingerprinting ? Gene therapy ? Mutation screening ? Drug discovery ? Classification of organisms ? Genotyping ? Molecular Archaeology ? Molecular Epidemiology ? Molecular Ecology ? Bioinformatics ? Genomic cloning ? Site-directed mutagenesis ? Gene expression studies
  • 16. Applications of PCR Molecular Identification Sequencing Genetic Engineering ? Molecular Archaeology ? Molecular Epidemiology ? Molecular Ecology ? DNA fingerprinting ? Classification of organisms ? Genotyping ? Pre-natal diagnosis ? Mutation screening ? Drug discovery ? Genetic matching ? Detection of pathogens ? Bioinformatics ? Genomic cloning ? Human Genome Project ? Site-directed mutagenesis ? Gene expression studies
  • 20. Detection of Unknown MutationsDetection of Unknown Mutations Molecular Identification:
  • 21. Classification of OrganismsClassification of Organisms 1) Relating to each other 2) Similarities 3) Differences * Fossils * Trace amounts * Small organisms ! DNA ! Molecular Identification: Insufficient data
  • 24. Detection Of PathogensDetection Of Pathogens Molecular Identification:
  • 26. SSEQUENCINGEQUENCING Nucleotides (dNTP) are modified (dideoxynucleotides = ddNTP) NO polymerisation after a dideoxynucleotide! Fragments of DNA differing only by one nucleotide are generated Nucleotides are either or
  • 28. SummarySummary blood, chorionic villus, amniotic fluid, semen, hair root, saliva 68,719,476,736 copies Gel Analysis, Restriction Digestion, Sequencing
  • 29. ConclusionConclusion The speedspeed and easeease of use, sensitivitysensitivity, specificityspecificity and robustnessrobustness of PCR has revolutionised molecular biology and made PCR the most widely used and powerful technique with great spectrum of research and diagnostic applications.