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Hawre Najmaddin
Hawre Najmaddin

Polymerase chain reaction (PCR) is a common technique used to amplify a specific segment of DNA. PCR works by using DNA polymerase to replicate the target DNA region through repeated heating and cooling cycles. The components of PCR include a DNA template, DNA polymerase, primers, nucleotides, and a buffer solution. The major steps of PCR are denaturation, annealing of primers, and extension. PCR has many applications in medicine, forensics, research, and genetics. The advantages are that it can detect pathogens that are difficult to culture and provide rapid results. Disadvantages include costs and potentially lower specificity compared to culture methods.

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Polymerase chain reaction (PCR) Prepared by Hawre najmaddin Zahra govand Omer fariq Blimat Kamran Bushra tarq Super visor :Dr. Ahmad Nawzad Hassan 1
Outline Introduction Principles Components Of PCR The steps of PCR Applications of PCR Advantages, disadvantage 2
Introduction Polymerase chain reaction (PCR) is a common laboratory technique used to make many copies (millions or billions!) of a particular region of DNA. This DNA region can be anything the experimenter is interested in. PCR is used in many areas of biology and medicine, including molecular biology research, medical diagnostics, and even some branches of ecology. 3
Principles Typically, the goal of PCR is to make enough of the target DNA region that it can be analyzed or used in some other way. For instance, DNA amplified by PCR may be sent for sequencing, visualized by gel electrophoresis, or cloned into a plasmid for further experiments. he PCR technique is based on the enzymatic replication of DNA. In PCR, a short segment of DNA is amplified using primer mediated enzymes. DNA Polymerase synthesises new strands of DNA complementary to the template DNA. The DNA polymerase can add a nucleotide to the pre-existing 3-OH group only. Therefore, a primer is required. Thus, more nucleotides are added to the 3 prime end of the DNA polymerase. 4
Components Of PCR Components Of PCR constitutes the following: 1.DNA Template The DNA of interest from the sample. 2.DNA Polymerase Taq Polymerase is used. It is thermostable and does not denature at very high temperatures. 3.Oligonucleotide Primers- These are the short stretches of single- stranded DNA complementary to the 3 ends of sense and anti-sense strands. 4.Deoxyribonucleotide triphosphate These provide energy for polymerization and are the building blocks for the synthesis of DNA. These are single units of bases. 5.Buffer System Magnesium and Potassium provide optimum conditions for DNA denaturation and renaturation. It is also important for fidelity, polymerase activity, and stability. 5
The steps of PCR PCR proceeds in THREE distinct steps Governed by Temperature: 6
Applications of PCR The following are the applications of PCR : Medicine Testing of genetic disease mutations. Monitoring the gene in gene therapy.Detecting disease-causing genes in the parents. Forensic Science Used as a tool in genetic fingerprinting. Identifying the criminal from millions of people.Paternity tests Research and Genetics Compare the genome of two organisms in genomic studies. In the phylogenetic analysis of DNA from any source such as fossils. Analysis of gene expression. Gene Mapping 7
Advantages, disadvantage Advantages of PCR Testing Valuable for detecting specific pathogens that are difficult to culture in vitro or require a long cultivation period Significantly more rapid in providing results compared to culturing o Enables earlier informed decision making o Rapid diagnosis of bacteremia, particularly for low levels of bacteria in specimens Useful in detecting cases in extra pulmonary specimens which may be missed by smear and/or culture Valuable screening tool o PCR is still considered an adjunct test for certain diagnostic tests that still rely on smear and culture, such as tuberculosis 8
disadvantage Supply costs, machinery fees, training expenses Potentially lower specificity compared to culture and staining Since specific primers are used to identify different microorganisms, physicians often need to list potential microorganisms before performing selective PCR 9

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medical ggyhyfgthgyhyhyujvvujubkikikikibkik

  • 1. Polymerase chain reaction (PCR) Prepared by Hawre najmaddin Zahra govand Omer fariq Blimat Kamran Bushra tarq Super visor :Dr. Ahmad Nawzad Hassan 1
  • 2. Outline Introduction Principles Components Of PCR The steps of PCR Applications of PCR Advantages, disadvantage 2
  • 3. Introduction Polymerase chain reaction (PCR) is a common laboratory technique used to make many copies (millions or billions!) of a particular region of DNA. This DNA region can be anything the experimenter is interested in. PCR is used in many areas of biology and medicine, including molecular biology research, medical diagnostics, and even some branches of ecology. 3
  • 4. Principles Typically, the goal of PCR is to make enough of the target DNA region that it can be analyzed or used in some other way. For instance, DNA amplified by PCR may be sent for sequencing, visualized by gel electrophoresis, or cloned into a plasmid for further experiments. he PCR technique is based on the enzymatic replication of DNA. In PCR, a short segment of DNA is amplified using primer mediated enzymes. DNA Polymerase synthesises new strands of DNA complementary to the template DNA. The DNA polymerase can add a nucleotide to the pre-existing 3-OH group only. Therefore, a primer is required. Thus, more nucleotides are added to the 3 prime end of the DNA polymerase. 4
  • 5. Components Of PCR Components Of PCR constitutes the following: 1.DNA Template The DNA of interest from the sample. 2.DNA Polymerase Taq Polymerase is used. It is thermostable and does not denature at very high temperatures. 3.Oligonucleotide Primers- These are the short stretches of single- stranded DNA complementary to the 3 ends of sense and anti-sense strands. 4.Deoxyribonucleotide triphosphate These provide energy for polymerization and are the building blocks for the synthesis of DNA. These are single units of bases. 5.Buffer System Magnesium and Potassium provide optimum conditions for DNA denaturation and renaturation. It is also important for fidelity, polymerase activity, and stability. 5
  • 6. The steps of PCR PCR proceeds in THREE distinct steps Governed by Temperature: 6
  • 7. Applications of PCR The following are the applications of PCR : Medicine Testing of genetic disease mutations. Monitoring the gene in gene therapy.Detecting disease-causing genes in the parents. Forensic Science Used as a tool in genetic fingerprinting. Identifying the criminal from millions of people.Paternity tests Research and Genetics Compare the genome of two organisms in genomic studies. In the phylogenetic analysis of DNA from any source such as fossils. Analysis of gene expression. Gene Mapping 7
  • 8. Advantages, disadvantage Advantages of PCR Testing Valuable for detecting specific pathogens that are difficult to culture in vitro or require a long cultivation period Significantly more rapid in providing results compared to culturing o Enables earlier informed decision making o Rapid diagnosis of bacteremia, particularly for low levels of bacteria in specimens Useful in detecting cases in extra pulmonary specimens which may be missed by smear and/or culture Valuable screening tool o PCR is still considered an adjunct test for certain diagnostic tests that still rely on smear and culture, such as tuberculosis 8
  • 9. disadvantage Supply costs, machinery fees, training expenses Potentially lower specificity compared to culture and staining Since specific primers are used to identify different microorganisms, physicians often need to list potential microorganisms before performing selective PCR 9