The polymerase chain reaction (PCR) is a technique used to amplify a specific region of DNA through a series of temperature changes and primer hybridization. It involves heating and cooling of the DNA sample in a series of cycles to denature the DNA, anneal primers, and extend new strands. This results in exponential amplification of the target DNA region, producing millions of copies. Key components of PCR include DNA template, primers, DNA polymerase, nucleotides, and buffer. It has numerous applications in molecular biology research and medical diagnostics.
PCR is a technique used to amplify a specific DNA sequence using thermal cycling. It involves denaturing double-stranded DNA into single strands, annealing primers to the DNA template, and extending the primers using a DNA polymerase to synthesize the complementary strand. This three-step cycle of denaturation, annealing and extension is repeated for typically 30-40 cycles to exponentially amplify the target DNA sequence.
Polymerase chain reaction (PCR) is a technique used to amplify a single copy of a DNA segment across orders of magnitude, generating thousands to millions of copies. PCR involves repeated cycles of heating and cooling of the DNA sample to denature and separate the DNA strands, followed by primer annealing and polymerase extension. This allows for exponential amplification of the target DNA sequence. PCR is commonly used in clinical and research applications such as disease diagnosis, genetic analysis, and forensic identification.
Polymerase chain reaction (PCR) is a laboratory technique for amplifying a specific DNA sequence. It involves repeated cycles of heating and cooling of the DNA sample to separate and copy the DNA strands. During each cycle, the DNA strands are separated by heating, primers anneal to the DNA by cooling, and the DNA polymerase enzyme synthesizes complementary DNA strands by extending the primers. This process results in exponential amplification of the target DNA sequence, generating millions of copies. PCR was invented by Kary Mullis in the 1980s and uses the thermostable Taq polymerase enzyme from bacteria. It has become essential to many areas of science including genetics, medicine and forensics.
PCR is a technique used to amplify specific DNA sequences. It involves repeated cycles of heating and cooling of the DNA sample to denature the double-stranded DNA, anneal primers, and extend new strands using a DNA polymerase. This process is repeated many times, exponentially amplifying the target DNA sequence. PCR is widely used in scientific research and forensic analysis.
The document discusses polymerase chain reaction (PCR), a technique used to amplify a single or few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. It explains that PCR uses DNA polymerase to replicate a specific DNA segment defined by a pair of primers that flank the region of interest. The key steps of PCR including denaturation of DNA, annealing of primers, and extension of primers by DNA polymerase are described. Common applications and requirements of PCR like thermostable DNA polymerase and temperature cycling are also summarized.
This is a powerpoint file of a practical class taken by Dr. Karthikeyan Pethsuamay for the first year MBBS students of AIIMS, New Delhi. Feel free to download and use for educational purposes. Happy learning and teaching!
Don't forget to watch the YouTube video.
The document discusses polymerase chain reaction (PCR), a technique used to amplify a single or few copies of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. It requires primers that flank the target DNA sequence, a heat-stable DNA polymerase, and repeated cycles of heating and cooling to denature and extend DNA strands. Key steps involve DNA denaturation, primer annealing, and polymerase extension. PCR is useful for applications like disease screening, forensics, genetic engineering and more due to its speed, sensitivity and ability to amplify small amounts of DNA.
This PPT shows the general information about PCR principles and gene expression analysis. It might be useful for researchers, students working in the field of molecular biology and genomics.
PCR is a technique used to amplify a specific DNA sequence using thermal cycling. It involves denaturing double-stranded DNA into single strands, annealing primers to the DNA template, and extending the primers using a DNA polymerase to synthesize the complementary strand. This three-step cycle of denaturation, annealing and extension is repeated for typically 30-40 cycles to exponentially amplify the target DNA sequence.
Polymerase chain reaction (PCR) is a technique used to amplify a single copy of a DNA segment across orders of magnitude, generating thousands to millions of copies. PCR involves repeated cycles of heating and cooling of the DNA sample to denature and separate the DNA strands, followed by primer annealing and polymerase extension. This allows for exponential amplification of the target DNA sequence. PCR is commonly used in clinical and research applications such as disease diagnosis, genetic analysis, and forensic identification.
Polymerase chain reaction (PCR) is a laboratory technique for amplifying a specific DNA sequence. It involves repeated cycles of heating and cooling of the DNA sample to separate and copy the DNA strands. During each cycle, the DNA strands are separated by heating, primers anneal to the DNA by cooling, and the DNA polymerase enzyme synthesizes complementary DNA strands by extending the primers. This process results in exponential amplification of the target DNA sequence, generating millions of copies. PCR was invented by Kary Mullis in the 1980s and uses the thermostable Taq polymerase enzyme from bacteria. It has become essential to many areas of science including genetics, medicine and forensics.
PCR is a technique used to amplify specific DNA sequences. It involves repeated cycles of heating and cooling of the DNA sample to denature the double-stranded DNA, anneal primers, and extend new strands using a DNA polymerase. This process is repeated many times, exponentially amplifying the target DNA sequence. PCR is widely used in scientific research and forensic analysis.
The document discusses polymerase chain reaction (PCR), a technique used to amplify a single or few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. It explains that PCR uses DNA polymerase to replicate a specific DNA segment defined by a pair of primers that flank the region of interest. The key steps of PCR including denaturation of DNA, annealing of primers, and extension of primers by DNA polymerase are described. Common applications and requirements of PCR like thermostable DNA polymerase and temperature cycling are also summarized.
This is a powerpoint file of a practical class taken by Dr. Karthikeyan Pethsuamay for the first year MBBS students of AIIMS, New Delhi. Feel free to download and use for educational purposes. Happy learning and teaching!
Don't forget to watch the YouTube video.
The document discusses polymerase chain reaction (PCR), a technique used to amplify a single or few copies of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. It requires primers that flank the target DNA sequence, a heat-stable DNA polymerase, and repeated cycles of heating and cooling to denature and extend DNA strands. Key steps involve DNA denaturation, primer annealing, and polymerase extension. PCR is useful for applications like disease screening, forensics, genetic engineering and more due to its speed, sensitivity and ability to amplify small amounts of DNA.
This PPT shows the general information about PCR principles and gene expression analysis. It might be useful for researchers, students working in the field of molecular biology and genomics.
Polymerase chain reaction is a technique used in molecular biology to amplify a single copy or a few copies of a segment of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence
The document discusses various techniques and applications of polymerase chain reaction (PCR). It describes the invention of PCR in 1982 and some key adaptations such as real-time PCR, which allows for quantitative analysis of DNA amplification in real time using fluorescent probes. Different types of PCR are also summarized, including reverse transcription PCR, nested PCR, multiplex PCR, and touchdown PCR. Components of real-time PCR and steps in the PCR process are outlined.
The Polymerase Chain Reaction (PCR) is a technique that allows for the amplification of specific DNA sequences. It involves cycling between heating and cooling steps to denature, anneal primers to, and extend DNA. This allows a small amount of DNA to be exponentially replicated, enabling applications like disease diagnosis, genetic identification, and DNA analysis. PCR requires DNA, primers, DNA polymerase, nucleotides, and thermal cycling to replicate the target DNA sequence.
PCR is a technique used to amplify specific DNA sequences. It involves repeated cycles of separating DNA strands through heating, annealing primers to the strands, and extending the strands with DNA polymerase. This process can produce billions of copies of the target DNA fragment. PCR is used in research, forensics, and medicine to detect genetic mutations and diseases.
This document provides information about polymerase chain reaction (PCR) and gel electrophoresis. It begins with an introduction to PCR, covering its history, basic procedure, requirements, applications and limitations. PCR is described as a technique for amplifying specific DNA sequences. The document then provides details on gel electrophoresis, including its use for analyzing amplified DNA from PCR. Gel electrophoresis separates DNA fragments by size when an electric current is applied through an agarose gel. Specific applications of both PCR and gel electrophoresis are given.
The document discusses various aspects of PCR including primer design, DNA polymerases used, different types of PCR such as multiplex PCR, nested PCR and real-time PCR. It also summarizes applications of PCR like cloning of PCR products, use in gene recombination and PCR-based mutagenesis. The key aspects covered are primer length and melting temperature for design, thermostability and fidelity improvements of DNA polymerases used in PCR and different modifications of standard PCR.
Nucleic acid amplification techniques involve synthesizing many copies of DNA or RNA from a template. They are classified into target amplification, probe amplification, and signal amplification. Polymerase chain reaction (PCR) is a commonly used target amplification technique that exponentially amplifies target DNA sequences in vitro using DNA polymerase. PCR has many applications including DNA sequencing, bacterial cloning, and detecting pathogens. It involves thermal cycling between denaturation, annealing of primers, and extension steps. Variations of PCR like real-time PCR, nested PCR, and digital PCR have further expanded its uses.
The polymerase chain reaction (PCR) is an in vitro technique used to amplify specific DNA sequences. It involves repeated cycles of denaturation, annealing of primers to the DNA template, and extension of the primers by DNA polymerase. The requirements for PCR include a targeted DNA or RNA sequence, primers complementary to regions flanking the target, PCR buffer, dNTPs, and a DNA polymerase such as Taq polymerase. PCR has various applications including parental diagnosis of genetic diseases, diagnosis of infections, DNA sequencing, gene expression studies, forensic analysis, and more. It has advantages such as simplicity, sensitivity, speed, and requiring small amounts of DNA, but also has disadvantages like risk of contamination and requirement of specialized equipment and skills.
Polymerase chain reaction (PCR) is a technique used to amplify a single copy of a DNA segment into millions of copies. PCR uses repeated cycles of heating and cooling of the DNA sample to denature and separate the double-stranded DNA, followed by annealing of primers and extension of the DNA strands by a DNA polymerase. This allows the targeted DNA segment to be exponentially amplified. A basic PCR setup requires DNA template, primers, dNTPs, buffer solution, bivalent cations such as magnesium, and a thermostable DNA polymerase like Taq polymerase. PCR has applications in cloning genes, detecting genetic mutations and microorganisms, and genetic fingerprinting.
Kary Banks is considered the great mind behind PCR. He developed PCR in 1985 while working at Cetus Corporation and was awarded the Nobel Prize in 1993. PCR allows for targeted amplification of specific DNA sequences, enabling their analysis even from very small samples. It involves heating and cooling of the DNA sample in the presence of primers, DNA polymerase, and nucleotides to exponentially amplify the target sequence. The amplified DNA can then be analyzed by gel electrophoresis.
The polymerase chain reaction (PCR) is a technique used to amplify a specific DNA sequence. It involves cycling between heating and cooling steps to denature and replicate DNA. The process results in exponential amplification of the target sequence. PCR requires a DNA template, primers, DNA polymerase, nucleotides, and buffer solutions. It goes through initialization, denaturation, annealing, and elongation steps in each cycle. PCR has many applications in medicine, research, forensics, and more.
this ppt contain about pcr technique and its three process,primers in pcr,dna polymerase in pcr,melting temp of dna in pcr and applications of pcr technology
Polymerase chain reaction (PCR) is a technique used to amplify a single copy of a DNA segment across orders of magnitude, generating thousands to millions of copies. PCR involves repeated cycles of heating and cooling of the DNA sample to denature and separate the DNA strands, followed by primer annealing and polymerase extension. This allows for exponential amplification of the target DNA segment. PCR is commonly used in clinical and research applications such as disease diagnosis, genetic analysis, and forensic identification.
Advances in instrumentation allow for more automated analysis. Automatic chemical analyzers can perform complex analyses quickly. The Coulter Counter uses electrical sensing to count and size particles like blood cells. Polymerase chain reaction (PCR) amplifies specific DNA sequences. It works in three stages: denaturing separates DNA strands, annealing attaches primers, and extending makes new strands with DNA polymerase. Automated blood culture instruments like the BACTEC system detect microbes in blood by monitoring changes in carbon dioxide or oxygen in sample vials over time. Positive readings indicate growth of microorganisms.
Polymerase chain reaction (PCR) allows for the amplification of specific DNA sequences. It is a sensitive, selective, and rapid technique that can amplify DNA from a single cell over 20-30 cycles. The PCR process involves strand separation, primer annealing, and polymerization through repeated heating and cooling cycles. PCR has many applications including disease diagnosis, cancer detection, forensics, and evolutionary studies. DNA sequencing determines the nucleotide sequence of genes and helps elucidate gene structure, expression, and function. Microarrays can analyze gene expression patterns across thousands of genes simultaneously and have applications in disease diagnosis and drug discovery.
The document discusses several types and applications of polymerase chain reaction (PCR). It begins by explaining the basic three-step cycling process of PCR: denaturation, annealing of primers, and extension. It then describes several variations of PCR including inverse PCR, anchored PCR, asymmetric PCR, real-time PCR (RT-PCR), and PCR for site-directed mutagenesis. Inverse PCR is used to amplify unknown flanking genomic regions, while anchored and asymmetric PCR are used to generate single-stranded DNA products for downstream applications like sequencing. RT-PCR amplifies RNA sequences by first generating cDNA. PCR mutagenesis introduces mutations through altered primer sequences.
Polymerase chain reaction (PCR) is a technique used to amplify small segments of DNA. During PCR, millions of copies of a DNA segment are made in just a few hours by separating the DNA strands, annealing primers, and extending new DNA strands using DNA polymerase. The key components of PCR are the target DNA, primers, DNA polymerase, nucleotides, and buffer solution. The PCR process involves denaturation, annealing, and extension steps that are repeated for multiple cycles to exponentially amplify the target DNA segment. PCR has many applications in basic research, applied research, and medical diagnosis.
Habib U Rahman presents on PCR (polymerase chain reaction), a technique used to amplify a specific segment of DNA. PCR involves three main steps: 1) denaturation to separate DNA strands at high temperature (94C for 1 minute), 2) annealing to attach primers at lower temperature (55C for 45 seconds), and 3) extension where Taq polymerase synthesizes new strands at 72C for 1-2 minutes. PCR is useful for replicating small amounts of DNA for further testing and analysis in molecular biology applications.
Polymerase chain reaction (PCR) is a technique used to amplify a specific DNA sequence. It involves repeated cycles of heating and cooling of the DNA sample to separate and copy the DNA strands. PCR requires DNA polymerase, primers, nucleotides, buffer, and thermal cycling. It has many applications including detecting pathogens, DNA fingerprinting, and genetic testing.
PCR is used to amplify specific DNA sequences. It involves repeated cycles of heating and cooling of the DNA sample to cause DNA replication between two primers that flank the target sequence. Each cycle doubles the amount of target DNA. After many cycles, the target is amplified exponentially into billions of copies. Key steps are denaturation to separate DNA strands, annealing to allow primers to bind, and extension to replicate the target using a DNA polymerase. Proper primer design is important for specificity of the amplification.
Polymerase chain reaction is a technique used in molecular biology to amplify a single copy or a few copies of a segment of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence
The document discusses various techniques and applications of polymerase chain reaction (PCR). It describes the invention of PCR in 1982 and some key adaptations such as real-time PCR, which allows for quantitative analysis of DNA amplification in real time using fluorescent probes. Different types of PCR are also summarized, including reverse transcription PCR, nested PCR, multiplex PCR, and touchdown PCR. Components of real-time PCR and steps in the PCR process are outlined.
The Polymerase Chain Reaction (PCR) is a technique that allows for the amplification of specific DNA sequences. It involves cycling between heating and cooling steps to denature, anneal primers to, and extend DNA. This allows a small amount of DNA to be exponentially replicated, enabling applications like disease diagnosis, genetic identification, and DNA analysis. PCR requires DNA, primers, DNA polymerase, nucleotides, and thermal cycling to replicate the target DNA sequence.
PCR is a technique used to amplify specific DNA sequences. It involves repeated cycles of separating DNA strands through heating, annealing primers to the strands, and extending the strands with DNA polymerase. This process can produce billions of copies of the target DNA fragment. PCR is used in research, forensics, and medicine to detect genetic mutations and diseases.
This document provides information about polymerase chain reaction (PCR) and gel electrophoresis. It begins with an introduction to PCR, covering its history, basic procedure, requirements, applications and limitations. PCR is described as a technique for amplifying specific DNA sequences. The document then provides details on gel electrophoresis, including its use for analyzing amplified DNA from PCR. Gel electrophoresis separates DNA fragments by size when an electric current is applied through an agarose gel. Specific applications of both PCR and gel electrophoresis are given.
The document discusses various aspects of PCR including primer design, DNA polymerases used, different types of PCR such as multiplex PCR, nested PCR and real-time PCR. It also summarizes applications of PCR like cloning of PCR products, use in gene recombination and PCR-based mutagenesis. The key aspects covered are primer length and melting temperature for design, thermostability and fidelity improvements of DNA polymerases used in PCR and different modifications of standard PCR.
Nucleic acid amplification techniques involve synthesizing many copies of DNA or RNA from a template. They are classified into target amplification, probe amplification, and signal amplification. Polymerase chain reaction (PCR) is a commonly used target amplification technique that exponentially amplifies target DNA sequences in vitro using DNA polymerase. PCR has many applications including DNA sequencing, bacterial cloning, and detecting pathogens. It involves thermal cycling between denaturation, annealing of primers, and extension steps. Variations of PCR like real-time PCR, nested PCR, and digital PCR have further expanded its uses.
The polymerase chain reaction (PCR) is an in vitro technique used to amplify specific DNA sequences. It involves repeated cycles of denaturation, annealing of primers to the DNA template, and extension of the primers by DNA polymerase. The requirements for PCR include a targeted DNA or RNA sequence, primers complementary to regions flanking the target, PCR buffer, dNTPs, and a DNA polymerase such as Taq polymerase. PCR has various applications including parental diagnosis of genetic diseases, diagnosis of infections, DNA sequencing, gene expression studies, forensic analysis, and more. It has advantages such as simplicity, sensitivity, speed, and requiring small amounts of DNA, but also has disadvantages like risk of contamination and requirement of specialized equipment and skills.
Polymerase chain reaction (PCR) is a technique used to amplify a single copy of a DNA segment into millions of copies. PCR uses repeated cycles of heating and cooling of the DNA sample to denature and separate the double-stranded DNA, followed by annealing of primers and extension of the DNA strands by a DNA polymerase. This allows the targeted DNA segment to be exponentially amplified. A basic PCR setup requires DNA template, primers, dNTPs, buffer solution, bivalent cations such as magnesium, and a thermostable DNA polymerase like Taq polymerase. PCR has applications in cloning genes, detecting genetic mutations and microorganisms, and genetic fingerprinting.
Kary Banks is considered the great mind behind PCR. He developed PCR in 1985 while working at Cetus Corporation and was awarded the Nobel Prize in 1993. PCR allows for targeted amplification of specific DNA sequences, enabling their analysis even from very small samples. It involves heating and cooling of the DNA sample in the presence of primers, DNA polymerase, and nucleotides to exponentially amplify the target sequence. The amplified DNA can then be analyzed by gel electrophoresis.
The polymerase chain reaction (PCR) is a technique used to amplify a specific DNA sequence. It involves cycling between heating and cooling steps to denature and replicate DNA. The process results in exponential amplification of the target sequence. PCR requires a DNA template, primers, DNA polymerase, nucleotides, and buffer solutions. It goes through initialization, denaturation, annealing, and elongation steps in each cycle. PCR has many applications in medicine, research, forensics, and more.
this ppt contain about pcr technique and its three process,primers in pcr,dna polymerase in pcr,melting temp of dna in pcr and applications of pcr technology
Polymerase chain reaction (PCR) is a technique used to amplify a single copy of a DNA segment across orders of magnitude, generating thousands to millions of copies. PCR involves repeated cycles of heating and cooling of the DNA sample to denature and separate the DNA strands, followed by primer annealing and polymerase extension. This allows for exponential amplification of the target DNA segment. PCR is commonly used in clinical and research applications such as disease diagnosis, genetic analysis, and forensic identification.
Advances in instrumentation allow for more automated analysis. Automatic chemical analyzers can perform complex analyses quickly. The Coulter Counter uses electrical sensing to count and size particles like blood cells. Polymerase chain reaction (PCR) amplifies specific DNA sequences. It works in three stages: denaturing separates DNA strands, annealing attaches primers, and extending makes new strands with DNA polymerase. Automated blood culture instruments like the BACTEC system detect microbes in blood by monitoring changes in carbon dioxide or oxygen in sample vials over time. Positive readings indicate growth of microorganisms.
Polymerase chain reaction (PCR) allows for the amplification of specific DNA sequences. It is a sensitive, selective, and rapid technique that can amplify DNA from a single cell over 20-30 cycles. The PCR process involves strand separation, primer annealing, and polymerization through repeated heating and cooling cycles. PCR has many applications including disease diagnosis, cancer detection, forensics, and evolutionary studies. DNA sequencing determines the nucleotide sequence of genes and helps elucidate gene structure, expression, and function. Microarrays can analyze gene expression patterns across thousands of genes simultaneously and have applications in disease diagnosis and drug discovery.
The document discusses several types and applications of polymerase chain reaction (PCR). It begins by explaining the basic three-step cycling process of PCR: denaturation, annealing of primers, and extension. It then describes several variations of PCR including inverse PCR, anchored PCR, asymmetric PCR, real-time PCR (RT-PCR), and PCR for site-directed mutagenesis. Inverse PCR is used to amplify unknown flanking genomic regions, while anchored and asymmetric PCR are used to generate single-stranded DNA products for downstream applications like sequencing. RT-PCR amplifies RNA sequences by first generating cDNA. PCR mutagenesis introduces mutations through altered primer sequences.
Polymerase chain reaction (PCR) is a technique used to amplify small segments of DNA. During PCR, millions of copies of a DNA segment are made in just a few hours by separating the DNA strands, annealing primers, and extending new DNA strands using DNA polymerase. The key components of PCR are the target DNA, primers, DNA polymerase, nucleotides, and buffer solution. The PCR process involves denaturation, annealing, and extension steps that are repeated for multiple cycles to exponentially amplify the target DNA segment. PCR has many applications in basic research, applied research, and medical diagnosis.
Habib U Rahman presents on PCR (polymerase chain reaction), a technique used to amplify a specific segment of DNA. PCR involves three main steps: 1) denaturation to separate DNA strands at high temperature (94C for 1 minute), 2) annealing to attach primers at lower temperature (55C for 45 seconds), and 3) extension where Taq polymerase synthesizes new strands at 72C for 1-2 minutes. PCR is useful for replicating small amounts of DNA for further testing and analysis in molecular biology applications.
Polymerase chain reaction (PCR) is a technique used to amplify a specific DNA sequence. It involves repeated cycles of heating and cooling of the DNA sample to separate and copy the DNA strands. PCR requires DNA polymerase, primers, nucleotides, buffer, and thermal cycling. It has many applications including detecting pathogens, DNA fingerprinting, and genetic testing.
PCR is used to amplify specific DNA sequences. It involves repeated cycles of heating and cooling of the DNA sample to cause DNA replication between two primers that flank the target sequence. Each cycle doubles the amount of target DNA. After many cycles, the target is amplified exponentially into billions of copies. Key steps are denaturation to separate DNA strands, annealing to allow primers to bind, and extension to replicate the target using a DNA polymerase. Proper primer design is important for specificity of the amplification.
PRIMER DESIGNING RECOMBINANT DNA TECHNOLOGY Primer designingnanamimomozano4562
The document discusses the process of polymerase chain reaction (PCR) and guidelines for designing efficient primers for PCR. PCR involves denaturation of DNA, annealing of primers to the single-stranded DNA, and extension of the primers by DNA polymerase. Successful PCR requires well-designed primers that are specific to the target sequence between 18-24 base pairs in length, have 40-60% GC content, and a melting temperature between 50-60°C. Primer design software such as Primer-BLAST can check for primer specificity and optimize parameters for primer design.
Polymerase chain reaction (PCR) is a technique used to amplify a specific region of DNA without cloning. It involves repeated cycles of separating the DNA strands by heating and synthesizing new strands with DNA polymerase. Two primers that flank the region of interest are used to determine the boundaries of the target sequence. During each cycle of PCR, the DNA is denatured by heating and the primers anneal to the single-stranded DNA. DNA polymerase then extends from the primers to synthesize new DNA strands. After multiple cycles, the target sequence is amplified exponentially into millions of copies. PCR is widely used in research, forensics, and medical diagnosis due to its simplicity, sensitivity, and ability to amplify specific DNA
PCR is a technique that amplifies a specific DNA sequence. It works by repeated cycles of heating and cooling of the DNA sample to separate, copy, and recombine strands. Each cycle approximately doubles the number of target sequences. This allows a very small initial sample to generate millions of copies of the target sequence. PCR is used in various applications including DNA sequencing, genetic disease diagnosis, cloning, and forensic analysis. It has become essential to many areas of biological research and medical diagnostics.
Primer design for PCR and analysis of gel picture Thoria Donia
The document discusses primer design for polymerase chain reaction (PCR). It explains that primers are short DNA sequences that are complementary to the target DNA segment to be amplified. The primers flank this segment. Effective primer design requires considering factors like primer length, GC content, melting temperature, specificity, and absence of repeats, hairpins and primer-dimer formation. Tools mentioned for primer design include Primer3, Primer3Plus, PrimerZ and PerlPrimer. The general steps involved in designing primers are obtaining the DNA sequence, setting parameters in primer design software, and checking the selected primer pairs for self-complementarity and specificity.
The document describes the polymerase chain reaction (PCR) process for amplifying a specific segment of DNA. PCR makes millions of copies of the target DNA segment in a few hours by using DNA polymerase enzyme and primers. It involves repeated cycles of heating and cooling of the DNA sample to denature and renature the DNA. The target DNA segment is amplified selectively between the forward and reverse primers. Key requirements for successful PCR include appropriate primer design and optimization of reaction conditions.
A detailed description about the basic steps involved in the - PCR - Polymerase Chain Reaction, its applications,its limitations and steps to overcome it.
A primer is a short nucleic acid sequence that provides a starting point for DNA synthesis during DNA replication. Primers are short strands of RNA synthesized by an enzyme called primase before DNA replication can occur. Arthur Kornberg discovered primers in 1956 while studying the mechanism of DNA replication. When designing primers for laboratory use, scientists consider factors like primer length, melting temperature, GC content, stability, and complementarity to avoid non-specific binding. Software is available to help design optimal primers for applications like PCR and DNA sequencing.
Polymerase chain reaction (PCR) is a technique developed by Kerry Mullis in 1984 that uses thermal cycling to amplify a specific DNA across several orders of magnitude, generating millions of copies of the target DNA segment. It involves repeated cycles of separating DNA strands through heating, annealing primers to the strands through cooling, and extending the primers with a thermostable DNA polymerase through heating. This allows for rapid and efficient amplification of targeted DNA regions.
This document discusses PCR (polymerase chain reaction), which is a technique that takes a specific sequence of DNA and amplifies it for further testing. It works by denaturing double-stranded DNA, annealing primers to the single strands, and using DNA polymerase to extend the primers, replicating the DNA. This process is repeated in cycles to exponentially amplify the target DNA sequence. The key components needed are DNA template, primers, thermostable DNA polymerase like Taq, and a thermal cycler machine to change temperatures for denaturation, annealing and extension steps. PCR has applications in molecular identification, genetic engineering, and DNA sequencing.
PCR involves copying a specific DNA segment through repeated cycles of heating and cooling. Each cycle consists of 3 stages - denaturation to separate DNA strands, annealing where primers attach to strands, and extension where new strands are synthesized. The process is carried out by a polymerase enzyme using primers, DNA bases, and thermal cycling to exponentially amplify the target DNA segment.
Polymerase chain reaction (PCR) is a technique used to amplify a specific region of DNA. It involves repeated cycles of heating and cooling of the DNA sample to separate the double-stranded DNA (denaturation), annealing primers to the single strands, and extending the primers with a DNA polymerase to replicate the DNA (extension). Over multiple cycles, this process exponentially amplifies the target DNA sequence. PCR requires a heat-stable DNA polymerase, primers, dNTPs, buffer, and a thermal cycler. It has many applications including disease diagnosis, forensics, genetic engineering, and molecular biology research.
Polymerase chain reaction (PCR) is a technique used to amplify a specific region of DNA. It involves repeated cycles of heating and cooling of the DNA sample in the presence of primers and a thermostable DNA polymerase. During each cycle, the DNA strand is separated from its complement at a high temperature, followed by lowering the temperature to allow primers to anneal and the polymerase to extend the primers to replicate the target DNA region. This process is repeated many times, exponentially amplifying the target DNA. PCR has many applications in research, forensics, medicine and molecular biology.
This document discusses polymerase chain reaction (PCR) and factors that affect it. It begins by defining PCR as a technique that amplifies a specific DNA sequence. It then outlines the basic requirements for PCR, including primers, thermostable DNA polymerase, and a thermal cycler. The document explains the three main steps in PCR: denaturation, annealing, and extension. It identifies several factors that can affect PCR efficiency and optimization, including primer length and sequence, target DNA length and sequence, annealing/extension temperatures and times, and reagent concentrations. The key factors for optimization are primer design, annealing temperature selection, and adjusting extension times and reagent concentrations.
Polymerase chain reaction (PCR) is a process that amplifies a specific DNA sequence using thermal cycling. It involves repeated cycles of heating and cooling of the DNA sample to cause DNA denaturation, primer annealing, and polymerase extension. PCR requires a DNA template, primers, DNA polymerase, dNTPs, and buffer. Multiple copies of the target DNA are generated in a thermal cycler through repetitive cycles of denaturation, annealing, and extension. Optimization of reaction components and cycling conditions is important for successful PCR amplification.
this section helps students how to prepare master mix solution and how to pcr. specially life life science fields such as biotechnology, biology, and medical laboratory
Polymerase chain reaction (PCR) is a laboratory technique used to amplify specific DNA sequences. It involves repeated cycles of heating and cooling of the DNA sample in the presence of primers and DNA polymerase. During each cycle, the DNA strands are separated by heating, then primers allow the polymerase to make copies of the target sequence. This results in exponential amplification, producing millions of copies of the target DNA. PCR is commonly used in research, forensics, medicine and many other applications to detect and analyze DNA.
The document provides an overview of polymerase chain reaction (PCR). It discusses the evolution of PCR since its invention in 1983. The key principles of PCR are that it uses thermal cycling to amplify a specific region of DNA across several orders of magnitude. Each cycle consists of DNA melting, primer annealing and strand extension steps. PCR requires DNA polymerase, primers, dNTPs and other reagents like buffers and divalent cations. The document discusses primer design considerations and different types of DNA polymerases and PCR techniques like reverse transcription PCR and real-time PCR.
Gene cloning and polymerase chain reaction Abhay jha
In these you are able to know about the gene cloning basic steps and Polymerase chain reaction process also there is an brief description about the ideal property shown by vectors which are lambda and M13 phases and there are lots of things in these slides
Influenza is caused by RNA viruses of the Orthomyxoviridae family, with three main types - A, B, and C. Type A is further divided into subtypes based on surface proteins hemagglutinin and neuraminidase, and can cause pandemics by antigenic shift or drift. The 1918 Spanish flu pandemic killed 20-50 million people. Later pandemics in 1957 and 1968 were caused by new subtype viruses. Seasonal flu is managed through vaccination against the predicted circulating strains each year.
This document provides information about extracting RNA and DNA from biological samples. It discusses the necessary equipment, the purpose of extraction, and the basic steps of the extraction process. For DNA extraction, the main steps involve lysing cells to expose the DNA, removing contaminants, and precipitating the purified DNA. For RNA extraction, similar steps are followed but special precautions must be taken due to RNA's instability. The document also notes common methods for RNA extraction and calculating concentrations of extracted RNA and DNA samples.
The document discusses influenza viruses, including their types (A, B, C), subtypes of type A (determined by surface proteins hemagglutinin and neuraminidase), and zoonotic potential. It covers topics like antigenic drift and shift, past pandemics, transmission and pathogenesis of influenza. The phases of pandemic influenza according to the WHO are outlined. High risk groups for severe illness from influenza are mentioned.
The polymerase chain reaction (PCR) is a technique used to amplify a specific region of DNA through a series of temperature changes and primer hybridization. The basic PCR protocol involves repeated cycles of denaturation to separate DNA strands, annealing of primers to the target sequences, and extension of the primers by a DNA polymerase. This results in exponential amplification of the target DNA region. PCR has numerous applications including DNA analysis, gene cloning, disease diagnosis, and genetic fingerprinting.
This document summarizes rapid detection methods for foodborne pathogen bacteria. It discusses how foodborne illnesses are a major public health problem and rapid detection of pathogens is needed. Several detection methods are outlined, including traditional culturing as well as newer techniques like PCR, real-time PCR, LAMP, and immunoassays. LAMP is highlighted as a new method that can rapidly detect pathogens under isothermal conditions. The document concludes that LAMP is a promising technique for pathogen detection due to its speed, simplicity and accuracy.
This document provides an outline for a presentation on HCV genotyping methods. It discusses the morphology and characteristics of HCV, its history and structure. It describes the six genotypes of HCV and their geographical distribution. It also discusses mutants of HCV, clinical importance of genotypes, laboratory diagnosis, and different methods for genotyping including direct sequencing, RFLP typing of the 5'UTR, and line probe assay (LiPA). The advantages of LiPA include it being easy, less time consuming and capable of reliably genotyping HCV RNA directly from clinical samples.
This document discusses genetic manipulation of carotenoid biosynthesis. It defines genetic engineering as the direct manipulation of an organism's genes using techniques like recombinant DNA and gene splicing. It explains that genetic manipulation can be used to alter existing species' characteristics or induce mutations to produce desirable traits. Specific techniques discussed include site-directed mutagenesis, protoplast fusion, and using shuttle and expression vectors. Carotenoids are described as pigments that protect plant structures and have health benefits when consumed by humans as antioxidants and for vitamin A activity. The locations and functions of carotenoid pigments are summarized.
Archaea are a domain of single-celled microorganisms that are distinct from bacteria and eukaryotes. They thrive in extreme environments like hot springs, salt lakes, and swamps. Archaea have unique cell structures and metabolisms that allow them to survive in these harsh conditions. They were originally classified as bacteria but are now recognized as a separate domain based on genetic and biochemical analysis. Archaea play important roles in biotechnology due to enzymes like Taq polymerase that are useful for PCR.
This document provides information on various microbiological culture media, including their uses, control organisms, incubation conditions, shelf life, and price. It describes 14 types of agar plates (blood agar, chocolate agar, MacConkey agar, etc.) and 10 types of broths/tubes (trypticase soy broth, triple sugar iron agar, motility indole lysine medium, etc.). For each, it lists the microorganisms it can be used to isolate or identify and how to incubate and store the medium.
This document provides an overview of biofilm formation in pathogens bacteria. It defines biofilms and describes their composition and structure. Biofilms provide bacteria advantages like increased antibiotic resistance. The document discusses where biofilms are commonly found and their role in various infectious diseases. It also reviews several studies examining biofilm formation in specific pathogens like Pseudomonas aeruginosa, Staphylococcus aureus, and Mycobacterium avium.
Antibodies are Y-shaped proteins produced by B cells that recognize and bind to foreign substances like viruses and bacteria. There are two main types: polyclonal antibodies which recognize multiple epitopes on an antigen and are produced through serum, and monoclonal antibodies which are derived from a single clone and recognize a single epitope. Monoclonal antibodies are important for research, diagnostics, and therapeutics. Antibody engineering techniques allow modifying antibodies to make them more effective, such as humanizing mouse antibodies to reduce immunogenicity.
The document discusses various types of viruses that can be used as biopesticides, including baculoviruses like nucleopolyhedrosis viruses and granulosis viruses which infect insect hosts and form crystalline protein structures allowing them to survive outside the host, as well as less classified group C baculoviruses and entomopox viruses. Baculoviruses like NPVs and GVs produce occlusion bodies within infected cells that protect the virus and allow it to be transmitted between hosts, making them useful for biocontrol.
This presentation outline summarizes information about the hepatitis C virus (HCV). It begins with the morphology, history, and structure of HCV. It then discusses the epidemiology and genotypes of the virus. The presentation also covers HCV mutants, methods for genotyping HCV including molecular techniques and RFLP analysis, and concludes with the clinical importance of HCV genotyping.
This document discusses enteric bacterial diseases caused by Escherichia coli, specifically enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli (EHEC). It provides background on the classification and epidemiology of EPEC, which is a leading cause of diarrhea in developing countries. The document then describes methods for the laboratory diagnosis of EPEC, including molecular techniques like PCR and sequencing to detect and identify these diarrheagenic E. coli strains.
Walmart Business+ and Spark Good for Nonprofits.pdfTechSoup
"Learn about all the ways Walmart supports nonprofit organizations.
You will hear from Liz Willett, the Head of Nonprofits, and hear about what Walmart is doing to help nonprofits, including Walmart Business and Spark Good. Walmart Business+ is a new offer for nonprofits that offers discounts and also streamlines nonprofits order and expense tracking, saving time and money.
The webinar may also give some examples on how nonprofits can best leverage Walmart Business+.
The event will cover the following::
Walmart Business + (https://business.walmart.com/plus) is a new shopping experience for nonprofits, schools, and local business customers that connects an exclusive online shopping experience to stores. Benefits include free delivery and shipping, a 'Spend Analytics” feature, special discounts, deals and tax-exempt shopping.
Special TechSoup offer for a free 180 days membership, and up to $150 in discounts on eligible orders.
Spark Good (walmart.com/sparkgood) is a charitable platform that enables nonprofits to receive donations directly from customers and associates.
Answers about how you can do more with Walmart!"
How to Make a Field Mandatory in Odoo 17Celine George
In Odoo, making a field required can be done through both Python code and XML views. When you set the required attribute to True in Python code, it makes the field required across all views where it's used. Conversely, when you set the required attribute in XML views, it makes the field required only in the context of that particular view.
How to Setup Warehouse & Location in Odoo 17 InventoryCeline George
In this slide, we'll explore how to set up warehouses and locations in Odoo 17 Inventory. This will help us manage our stock effectively, track inventory levels, and streamline warehouse operations.
Communicating effectively and consistently with students can help them feel at ease during their learning experience and provide the instructor with a communication trail to track the course's progress. This workshop will take you through constructing an engaging course container to facilitate effective communication.
it describes the bony anatomy including the femoral head , acetabulum, labrum . also discusses the capsule , ligaments . muscle that act on the hip joint and the range of motion are outlined. factors affecting hip joint stability and weight transmission through the joint are summarized.
Philippine Edukasyong Pantahanan at Pangkabuhayan (EPP) CurriculumMJDuyan
(𝐓𝐋𝐄 𝟏𝟎𝟎) (𝐋𝐞𝐬𝐬𝐨𝐧 𝟏)-𝐏𝐫𝐞𝐥𝐢𝐦𝐬
𝐃𝐢𝐬𝐜𝐮𝐬𝐬 𝐭𝐡𝐞 𝐄𝐏𝐏 𝐂𝐮𝐫𝐫𝐢𝐜𝐮𝐥𝐮𝐦 𝐢𝐧 𝐭𝐡𝐞 𝐏𝐡𝐢𝐥𝐢𝐩𝐩𝐢𝐧𝐞𝐬:
- Understand the goals and objectives of the Edukasyong Pantahanan at Pangkabuhayan (EPP) curriculum, recognizing its importance in fostering practical life skills and values among students. Students will also be able to identify the key components and subjects covered, such as agriculture, home economics, industrial arts, and information and communication technology.
𝐄𝐱𝐩𝐥𝐚𝐢𝐧 𝐭𝐡𝐞 𝐍𝐚𝐭𝐮𝐫𝐞 𝐚𝐧𝐝 𝐒𝐜𝐨𝐩𝐞 𝐨𝐟 𝐚𝐧 𝐄𝐧𝐭𝐫𝐞𝐩𝐫𝐞𝐧𝐞𝐮𝐫:
-Define entrepreneurship, distinguishing it from general business activities by emphasizing its focus on innovation, risk-taking, and value creation. Students will describe the characteristics and traits of successful entrepreneurs, including their roles and responsibilities, and discuss the broader economic and social impacts of entrepreneurial activities on both local and global scales.
LAND USE LAND COVER AND NDVI OF MIRZAPUR DISTRICT, UPRAHUL
This Dissertation explores the particular circumstances of Mirzapur, a region located in the
core of India. Mirzapur, with its varied terrains and abundant biodiversity, offers an optimal
environment for investigating the changes in vegetation cover dynamics. Our study utilizes
advanced technologies such as GIS (Geographic Information Systems) and Remote sensing to
analyze the transformations that have taken place over the course of a decade.
The complex relationship between human activities and the environment has been the focus
of extensive research and worry. As the global community grapples with swift urbanization,
population expansion, and economic progress, the effects on natural ecosystems are becoming
more evident. A crucial element of this impact is the alteration of vegetation cover, which plays a
significant role in maintaining the ecological equilibrium of our planet.Land serves as the foundation for all human activities and provides the necessary materials for
these activities. As the most crucial natural resource, its utilization by humans results in different
'Land uses,' which are determined by both human activities and the physical characteristics of the
land.
The utilization of land is impacted by human needs and environmental factors. In countries
like India, rapid population growth and the emphasis on extensive resource exploitation can lead
to significant land degradation, adversely affecting the region's land cover.
Therefore, human intervention has significantly influenced land use patterns over many
centuries, evolving its structure over time and space. In the present era, these changes have
accelerated due to factors such as agriculture and urbanization. Information regarding land use and
cover is essential for various planning and management tasks related to the Earth's surface,
providing crucial environmental data for scientific, resource management, policy purposes, and
diverse human activities.
Accurate understanding of land use and cover is imperative for the development planning
of any area. Consequently, a wide range of professionals, including earth system scientists, land
and water managers, and urban planners, are interested in obtaining data on land use and cover
changes, conversion trends, and other related patterns. The spatial dimensions of land use and
cover support policymakers and scientists in making well-informed decisions, as alterations in
these patterns indicate shifts in economic and social conditions. Monitoring such changes with the
help of Advanced technologies like Remote Sensing and Geographic Information Systems is
crucial for coordinated efforts across different administrative levels. Advanced technologies like
Remote Sensing and Geographic Information Systems
9
Changes in vegetation cover refer to variations in the distribution, composition, and overall
structure of plant communities across different temporal and spatial scales. These changes can
occur natural.
4. History
One of the most powerful tools in
molecular biology
Invented by Kary Mullis in 1983,
resulting in his Nobel Prize in
Chemistry
First published account appeared
in 1985.
Awarded Nobel Prize for
Chemistry in 1993.
4/28/2012 4
5. The reaction mixture
1. DNA (purified or a crude extract)
2. Primers specific for the target DNA
3. Free nucleotides (A, G, T, C)
4. DNA polymerase
5. Buffer (containing magnesium)
6. The reaction mixture
1- DNA template that contains the DNA region (target) to be
amplified.
2- One or more primers, which are complementary to the
DNA regions at the 5' (five prime) and 3' (three prime)
ends of the DNA region.
3- A DNA polymerase such as Taq polymerase or another
DNA polymerase with a temperature optimum at around
70 C.
4/28/2012 6
7. The reaction mixture
, (dNTPs) from which the
DNA polymerase builds the new DNA
, which provides a suitable chemical environment for
the DNA Polymerase
, or ions; generally
Mg2+ is used, but Mn2+ can be utilized for PCR-mediated
DNA mutagenesis, as higher Mn2+ concentration increases
the error rate during DNA synthesis
Monovalent cation ions.
4/28/2012 7
8. Primers
• On the other hand, the length of a primer is limited by:
the maximum temperature allowed to be applied in order to melt it,
as increases with the of the primer
that are too high, i.e., above , can cause problems:
since the is at such temperatures
• The of a primer is generally from 15 to 40 ,
with a between
calculating:
Tm =4(G+C)+2(A+T)
Software
9. Primers
• The DNA fragment to be amplified is determined by
selecting primers
• Primers are :
short, artificial DNA strands
often not more than 50 and usually only 18 to 25 base
pairs long
that are complementary to the beginning or the end of the
DNA fragment to be amplified
• They anneal by adhering to the DNA template at these
starting & ending points,
where the DNA polymerase binds and begins the
synthesis of the new DNA strand
10. Primer 3' terminus
• Primer 3' terminus design is critical to PCR success
since the primer extends from the 3' end
• The 3' end should not be complementary over greater
than 3-4 bases to any region of the other primer
(or even the same primer) used in the reaction
and must provide correct base matching to the
template
• There are computer programs to help design primers
Genrunner
13. The basic protocol
1. Denaturation of DNA to single
strands
2. Annealing of primers to DNA
3. Extension by polymerase
4. Repeat 30-35 times
14. Procedure
The PCR process usually consists of 20 -35
each cycle consists of :
1. The has to be heated to (or 98 C if extremely thermostable
polymerases are used)
in order to separate the strands
This step is called denaturing:
it breaks apart the that connect the two DNA strands
Prior to the first cycle:
the DNA is often denatured for an to ensure that the
and the ,
have completely separated and are now
usually , but up to minutes
Also certain polymerases at this step (
15. Procedure
2. After separating the DNA strands, the temperature is
so :
the primers can themselves to the
the temperature of this stage on the and is
usually their Tm (45-60 C)
A wrong temperature during the annealing step can result in :
primers not binding to the template DNA at all
or binding at random
Time: 1-2 minutes
16. Procedure
3. Finally, the DNA polymerase has to copy the DNA strands
It starts at the annealed primer and works its way along the DNA strand
this step is called elongation
the elongation temperature depends on the DNA polymerase:
Taq polymerase elongates optimally at a temperature of 72º C
• The time for this step depends:
1. both on the DNA polymerase itself
2. and on the length of the DNA fragment to be amplified
as a rule-of-thumb, this step takes 1 minute per 1000 bp
• A final elongation step is frequently used after the last cycle
to ensure that any remaining single stranded DNA is completely copied,
this differs from all other elongation steps, only in that it is longer,
typically 10-15 minutes
17. Primers
forward
5’ 3’
Target DNA
3’ 5’
reverse
26. One One billion in about 2 hours!
• At the end of each cycle, the amount of DNA
has doubled
• By the end of 30 cycles, you will have about 1
billion molecules from the original one you
started with!!
230=1,073,741,824
27. The basic protocol—what’s in the tube
5’ 3’
Target DNA
3’ 5’
A
B Free
primers nucleotides
Mg2+ Mg
2+
Buffer
Taq DNA Mg2+ containing
Mg2+
polymerase Mg2+ magnesium
Mg2+
28. 4/28/2012 Free Template from www.brainybetty.com 28
29. Optimising the PCR Reaction
C G Denaturation -
- Annealing -
Primer extension -
--
Ramp -
dNTP -
DNA Template DNA -
PCR -
-
Tm -
30. PCR optimization
1. For the preparation of reaction mixture, a laminar flow
cabinet with UV lamp is recommended
2. Fresh gloves should be used for each PCR step
3. As well as displacement pipettes with aerosol filters
4. The reagents for PCR should be prepared separately and used
solely for this purpose
5. Aliquots should be stored separately from other DNA
samples
6. A control reaction (inner control), omitting template
DNA, should always be performed, to confirm :
a. the absence of contamination
b. or primer multimer formation
31. Applications of PCR
1. the detection of hereditary diseases
2. the identification of genetic fingerprints
3. the diagnosis of infectious diseases
4. the cloning of genes
5. paternity testing
6. and DNA computing
32.
33. How It Works
• Heating/cooling
• Capillary surface area
Intake
• Single chamber
– holds 32 capillaries
• Photohybrids measure
fluorescence at 530,
640 and 705nm
41. The use of multiple, unique primer sets within a single PCR reaction ,
to produce amplicons of varying sizes specific to different DNA sequences
• By targeting multiple genes at once,
additional information may be elicited from a single test run that otherwise
:
would require several times the reagents and technician time to perform
• Annealing temperatures for each of the primer sets ,
must be optimized to work correctly within a single reaction
and amplicon sizes should be separated by enough difference,
in final base pair length to form distinct bands via gel electrophoresis
42. Multiplex PCR
• PCR reactions can be devised in which several
targets are amplified simultaneously often used
in diagnostic applications.
44. Nested PCR
is intended to reduce the contaminations in
products due to the amplification of
unexpected primer binding sites
• Two sets of primers are used in two successive
PCR runs
the second set intended to amplify a secondary
target within the first run product
• This is very successful, but requires more
detailed knowledge of the sequences involved
45. RT-PCR
RT-PCR (Reverse Transcription PCR) is the method
used to amplify, isolate or identify a known sequence
from a cell or tissues RNA library
• Essentially normal PCR preceded by transcription by
Reverse transcriptase (to convert the RNA to cDNA)
this is widely used in :
1. expression mapping, determining when and where
certain genes are expressed
2. detection of RNA viruses
46. Colony PCR
Bacterial clones (E.coli) can be screened for
the correct ligation products
• Selected colonies are picked with a sterile
toothpick from an agarose plate,
and dabbed into the master mix or sterile
water,
primers (and the master mix) are added
the PCR protocol has to be started with an
extended time at 95ºC
48. Uses of PCR
PCR can be used for a broad variety of experiments and analyses:
1. Genetic fingerprinting
• is a forensic technique used to identify a person by comparing his or
her DNA with a given sample
• An example is blood from a crime scene being genetically compared
to blood from a suspect
• The sample may contain only a tiny amount of DNA ,
(obtained from a source such as blood, semen, saliva, hair, or other
organic material)
49. Uses of PCR
2. Detection of hereditary diseases
• The detection of hereditary diseases in a given genome is a
long and difficult process, :
which can be shortened significantly by using PCR
• Each gene in question can easily be amplified through PCR by
using the appropriate primers :
and then sequenced to detect mutations
50. Uses of PCR
3. Viral diseases
• can be detected using PCR through amplification of
the viral DNA
• This analysis is possible right after infection,
which can be from several days to several months
before actual symptoms occur
• Such early diagnoses give physicians a significant
lead in treatment
• Treatment evaluation, viral load
• Genotyping, viruses, bacteria
51. Uses of PCR
4. Mutagenesis
• Mutations can be introduced into copied DNA sequences,
in two fundamentally different ways in the PCR process
• Site-directed mutagenesis allows the experimenter to introduce a mutation
at a specific location on the DNA strand
• Usually, the desired mutation is incorporated in the primers used for the
PCR program
• Random mutagenesis, is based on the use of error-prone polymerases in the
PCR process
the location and nature of the mutations cannot be controlled
• One application of random mutagenesis is :
to analyze structure-function relationships of a protein
• By randomly altering a DNA sequence:
one can compare the resulting protein ,
with the original and determine the function of each part of the protein
52. Uses of PCR
• 5. Genotyping of specific mutations
• Through the use of allele-specific PCR,
one can easily determine which allele of a mutation or
polymorphism an individual has
• Here, one of the two primers is common,
and would anneal a short distance away from the mutation,
while the other anneals right on the variation
• The 3' end of the allele-specific primer is modified,
to only anneal if it matches one of the alleles