Polymerase chain reaction (PCR) is a technique used to amplify a single or few copies of a DNA sequence to generate thousands to millions of copies. It involves repeating cycles of denaturing DNA, annealing primers to the single strands, and extending the primers with a DNA polymerase. Real-time PCR allows quantification of the PCR product at each cycle by detecting fluorescence from DNA-binding dyes or probe hydrolysis. It has applications in diagnosing diseases, detecting gene expression, identifying pathogens, and assessing genetically modified organisms.
The advent of the polymerase chain reaction (PCR) radically transformed biological science from the time it was first discovered (Mullis, 1990). For the first time, it allowed for specific detection and production of large amounts of DNA. PCR-based strategies have propelled huge scientific endeavors such as the Human Genome Project. The technique is currently widely used by clinicians and researchers to diagnose diseases, clone and sequence genes, and carry out sophisticated quantitative and genomic studies in a rapid and very sensitive manner. One of the most important medical applications of the classical PCR method is the detection of pathogens. In addition, the PCR assay is used in forensic medicine to identify criminals. Because of its widespread use, it is important to understand the basic principles of PCR and how its use can be modified to provide for sophisticated analysis of genes and the genome
PCR,polymerase chain reaction.Basic concept of PCR.naveed ul mushtaq
PCR.Basic concept of PCR. Steps in PCR.
Quantitative real time polymerase chain reaction.Fluorescent dyes and probes.
Advantages real-time PCR.
Real-time PCR primer
Primer design software
The advent of the polymerase chain reaction (PCR) radically transformed biological science from the time it was first discovered (Mullis, 1990). For the first time, it allowed for specific detection and production of large amounts of DNA. PCR-based strategies have propelled huge scientific endeavors such as the Human Genome Project. The technique is currently widely used by clinicians and researchers to diagnose diseases, clone and sequence genes, and carry out sophisticated quantitative and genomic studies in a rapid and very sensitive manner. One of the most important medical applications of the classical PCR method is the detection of pathogens. In addition, the PCR assay is used in forensic medicine to identify criminals. Because of its widespread use, it is important to understand the basic principles of PCR and how its use can be modified to provide for sophisticated analysis of genes and the genome
PCR,polymerase chain reaction.Basic concept of PCR.naveed ul mushtaq
PCR.Basic concept of PCR. Steps in PCR.
Quantitative real time polymerase chain reaction.Fluorescent dyes and probes.
Advantages real-time PCR.
Real-time PCR primer
Primer design software
Real Time Polymerase Chain Reaction
Basics of Real Time PCR
Definition
Advantages
Principles
Instruments (Thermal Cyclers)
Useful terms
Real Time PCR Chemistry
Fluorescence Dyes
SYBR Green
EvaGreen
Melt Doctor
Fluorescence Probes
TaqMan Probe
Molecular Beacons
Scorpion Primers
SYBR Green In details
qPCR Set-Up
Assay Design
Data Analysis
Troubleshooting
PCR is a technique which is used to amplify the number of copies of a specific region of DNA, in order to produce enough DNA to be adequately tested.
Cell-free amplification for synthesizing multiple identical copies (billions) of any DNA of interest.
Basic tool for the molecular biologist.
The purpose of a PCR is to make a huge number of copies of a gene. As a result, it now becomes possible to analyze and characterize DNA fragments found in minute quantities in places like a drop of blood at a crime scene or a cell from an extinct dinosaur.
Like Xerox machine for gene copying.
The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2−ΔΔCT method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2−ΔΔCT method. In addition, we present the derivation and applications of two variations of the 2−ΔΔCT method that may be useful in the analysis of real-time, quantitative PCR data.
Introduction to Real Time PCR (Q-PCR/qPCR/qrt-PCR): qPCR Technology Webinar S...QIAGEN
This slidedeck introduces the concepts of real-time PCR and how to conduct a real-time PCR assay. The topics that are covered include an overview of real-time PCR chemistries, protocols, quantification methods, real-time PCR applications and factors for success.
RT-PCR (reverse transcription-polymerase chain reaction) is a variant of the polymerase chain reaction (PCR) which are now widely used. Traditionally RT-PCR involves two steps: the RT reaction and PCR amplification. RNA is first reverse transcribed into cDNA using a reverse transcriptase as described here, the resulting cDNA is used as templates for subsequent PCR amplification using primers specific for one or more genes. RT-PCR can be used to quantify mRNA levels from much smaller samples. In fact, this technique is sensitive enough to enable quantitation of RNA from a single cell.
REAL TIME PCR, principle of real time pcr, method for detection real time pcr, taq man probe, molecular beacons. application of real time pcr. difference between real time pcr and conventional pcr.
Real Time Polymerase Chain Reaction
Basics of Real Time PCR
Definition
Advantages
Principles
Instruments (Thermal Cyclers)
Useful terms
Real Time PCR Chemistry
Fluorescence Dyes
SYBR Green
EvaGreen
Melt Doctor
Fluorescence Probes
TaqMan Probe
Molecular Beacons
Scorpion Primers
SYBR Green In details
qPCR Set-Up
Assay Design
Data Analysis
Troubleshooting
PCR is a technique which is used to amplify the number of copies of a specific region of DNA, in order to produce enough DNA to be adequately tested.
Cell-free amplification for synthesizing multiple identical copies (billions) of any DNA of interest.
Basic tool for the molecular biologist.
The purpose of a PCR is to make a huge number of copies of a gene. As a result, it now becomes possible to analyze and characterize DNA fragments found in minute quantities in places like a drop of blood at a crime scene or a cell from an extinct dinosaur.
Like Xerox machine for gene copying.
The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2−ΔΔCT method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2−ΔΔCT method. In addition, we present the derivation and applications of two variations of the 2−ΔΔCT method that may be useful in the analysis of real-time, quantitative PCR data.
Introduction to Real Time PCR (Q-PCR/qPCR/qrt-PCR): qPCR Technology Webinar S...QIAGEN
This slidedeck introduces the concepts of real-time PCR and how to conduct a real-time PCR assay. The topics that are covered include an overview of real-time PCR chemistries, protocols, quantification methods, real-time PCR applications and factors for success.
RT-PCR (reverse transcription-polymerase chain reaction) is a variant of the polymerase chain reaction (PCR) which are now widely used. Traditionally RT-PCR involves two steps: the RT reaction and PCR amplification. RNA is first reverse transcribed into cDNA using a reverse transcriptase as described here, the resulting cDNA is used as templates for subsequent PCR amplification using primers specific for one or more genes. RT-PCR can be used to quantify mRNA levels from much smaller samples. In fact, this technique is sensitive enough to enable quantitation of RNA from a single cell.
REAL TIME PCR, principle of real time pcr, method for detection real time pcr, taq man probe, molecular beacons. application of real time pcr. difference between real time pcr and conventional pcr.
Basic Molecular Biology:
Molecular biology is the branch of biology that focuses on understanding the fundamental processes and mechanisms underlying life at the molecular level. It involves the study of biological molecules such as DNA, RNA, and proteins, and how they interact to regulate various cellular processes. Molecular biology techniques enable scientists to investigate genetic information, gene expression, and the structure and function of macromolecules.
Polymerase Chain Reaction (PCR):
Polymerase Chain Reaction (PCR) is a powerful molecular biology technique used to amplify and replicate a specific segment of DNA in a laboratory setting. PCR allows scientists to make millions of copies of a target DNA sequence in a short period. It consists of repeated cycles of denaturation (separation of DNA strands), annealing (binding of short DNA primers to the target sequence), and extension (synthesis of new DNA strands using a heat-stable DNA polymerase enzyme). PCR has diverse applications, including DNA sequencing, genetic testing, forensics, and the study of gene expression.
Reverse Transcription Polymerase Chain Reaction (RT-PCR):
Reverse Transcription Polymerase Chain Reaction (RT-PCR) is a variation of the standard PCR technique that is specifically used to amplify RNA molecules. It involves a two-step process. First, the RNA is reverse transcribed into complementary DNA (cDNA) using the enzyme reverse transcriptase. Then, the cDNA is amplified using standard PCR. RT-PCR is essential for studying gene expression, viral RNA detection (e.g., for diagnosing diseases like COVID-19), and a range of other applications where RNA analysis is crucial.
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
Essentials of Automations: Optimizing FME Workflows with ParametersSafe Software
Are you looking to streamline your workflows and boost your projects’ efficiency? Do you find yourself searching for ways to add flexibility and control over your FME workflows? If so, you’re in the right place.
Join us for an insightful dive into the world of FME parameters, a critical element in optimizing workflow efficiency. This webinar marks the beginning of our three-part “Essentials of Automation” series. This first webinar is designed to equip you with the knowledge and skills to utilize parameters effectively: enhancing the flexibility, maintainability, and user control of your FME projects.
Here’s what you’ll gain:
- Essentials of FME Parameters: Understand the pivotal role of parameters, including Reader/Writer, Transformer, User, and FME Flow categories. Discover how they are the key to unlocking automation and optimization within your workflows.
- Practical Applications in FME Form: Delve into key user parameter types including choice, connections, and file URLs. Allow users to control how a workflow runs, making your workflows more reusable. Learn to import values and deliver the best user experience for your workflows while enhancing accuracy.
- Optimization Strategies in FME Flow: Explore the creation and strategic deployment of parameters in FME Flow, including the use of deployment and geometry parameters, to maximize workflow efficiency.
- Pro Tips for Success: Gain insights on parameterizing connections and leveraging new features like Conditional Visibility for clarity and simplicity.
We’ll wrap up with a glimpse into future webinars, followed by a Q&A session to address your specific questions surrounding this topic.
Don’t miss this opportunity to elevate your FME expertise and drive your projects to new heights of efficiency.
Software Delivery At the Speed of AI: Inflectra Invests In AI-Powered QualityInflectra
In this insightful webinar, Inflectra explores how artificial intelligence (AI) is transforming software development and testing. Discover how AI-powered tools are revolutionizing every stage of the software development lifecycle (SDLC), from design and prototyping to testing, deployment, and monitoring.
Learn about:
• The Future of Testing: How AI is shifting testing towards verification, analysis, and higher-level skills, while reducing repetitive tasks.
• Test Automation: How AI-powered test case generation, optimization, and self-healing tests are making testing more efficient and effective.
• Visual Testing: Explore the emerging capabilities of AI in visual testing and how it's set to revolutionize UI verification.
• Inflectra's AI Solutions: See demonstrations of Inflectra's cutting-edge AI tools like the ChatGPT plugin and Azure Open AI platform, designed to streamline your testing process.
Whether you're a developer, tester, or QA professional, this webinar will give you valuable insights into how AI is shaping the future of software delivery.
JMeter webinar - integration with InfluxDB and GrafanaRTTS
Watch this recorded webinar about real-time monitoring of application performance. See how to integrate Apache JMeter, the open-source leader in performance testing, with InfluxDB, the open-source time-series database, and Grafana, the open-source analytics and visualization application.
In this webinar, we will review the benefits of leveraging InfluxDB and Grafana when executing load tests and demonstrate how these tools are used to visualize performance metrics.
Length: 30 minutes
Session Overview
-------------------------------------------
During this webinar, we will cover the following topics while demonstrating the integrations of JMeter, InfluxDB and Grafana:
- What out-of-the-box solutions are available for real-time monitoring JMeter tests?
- What are the benefits of integrating InfluxDB and Grafana into the load testing stack?
- Which features are provided by Grafana?
- Demonstration of InfluxDB and Grafana using a practice web application
To view the webinar recording, go to:
https://www.rttsweb.com/jmeter-integration-webinar
Accelerate your Kubernetes clusters with Varnish CachingThijs Feryn
A presentation about the usage and availability of Varnish on Kubernetes. This talk explores the capabilities of Varnish caching and shows how to use the Varnish Helm chart to deploy it to Kubernetes.
This presentation was delivered at K8SUG Singapore. See https://feryn.eu/presentations/accelerate-your-kubernetes-clusters-with-varnish-caching-k8sug-singapore-28-2024 for more details.
Transcript: Selling digital books in 2024: Insights from industry leaders - T...BookNet Canada
The publishing industry has been selling digital audiobooks and ebooks for over a decade and has found its groove. What’s changed? What has stayed the same? Where do we go from here? Join a group of leading sales peers from across the industry for a conversation about the lessons learned since the popularization of digital books, best practices, digital book supply chain management, and more.
Link to video recording: https://bnctechforum.ca/sessions/selling-digital-books-in-2024-insights-from-industry-leaders/
Presented by BookNet Canada on May 28, 2024, with support from the Department of Canadian Heritage.
Elevating Tactical DDD Patterns Through Object CalisthenicsDorra BARTAGUIZ
After immersing yourself in the blue book and its red counterpart, attending DDD-focused conferences, and applying tactical patterns, you're left with a crucial question: How do I ensure my design is effective? Tactical patterns within Domain-Driven Design (DDD) serve as guiding principles for creating clear and manageable domain models. However, achieving success with these patterns requires additional guidance. Interestingly, we've observed that a set of constraints initially designed for training purposes remarkably aligns with effective pattern implementation, offering a more ‘mechanical’ approach. Let's explore together how Object Calisthenics can elevate the design of your tactical DDD patterns, offering concrete help for those venturing into DDD for the first time!
Securing your Kubernetes cluster_ a step-by-step guide to success !KatiaHIMEUR1
Today, after several years of existence, an extremely active community and an ultra-dynamic ecosystem, Kubernetes has established itself as the de facto standard in container orchestration. Thanks to a wide range of managed services, it has never been so easy to set up a ready-to-use Kubernetes cluster.
However, this ease of use means that the subject of security in Kubernetes is often left for later, or even neglected. This exposes companies to significant risks.
In this talk, I'll show you step-by-step how to secure your Kubernetes cluster for greater peace of mind and reliability.
The Art of the Pitch: WordPress Relationships and SalesLaura Byrne
Clients don’t know what they don’t know. What web solutions are right for them? How does WordPress come into the picture? How do you make sure you understand scope and timeline? What do you do if sometime changes?
All these questions and more will be explored as we talk about matching clients’ needs with what your agency offers without pulling teeth or pulling your hair out. Practical tips, and strategies for successful relationship building that leads to closing the deal.
DevOps and Testing slides at DASA ConnectKari Kakkonen
My and Rik Marselis slides at 30.5.2024 DASA Connect conference. We discuss about what is testing, then what is agile testing and finally what is Testing in DevOps. Finally we had lovely workshop with the participants trying to find out different ways to think about quality and testing in different parts of the DevOps infinity loop.
UiPath Test Automation using UiPath Test Suite series, part 4DianaGray10
Welcome to UiPath Test Automation using UiPath Test Suite series part 4. In this session, we will cover Test Manager overview along with SAP heatmap.
The UiPath Test Manager overview with SAP heatmap webinar offers a concise yet comprehensive exploration of the role of a Test Manager within SAP environments, coupled with the utilization of heatmaps for effective testing strategies.
Participants will gain insights into the responsibilities, challenges, and best practices associated with test management in SAP projects. Additionally, the webinar delves into the significance of heatmaps as a visual aid for identifying testing priorities, areas of risk, and resource allocation within SAP landscapes. Through this session, attendees can expect to enhance their understanding of test management principles while learning practical approaches to optimize testing processes in SAP environments using heatmap visualization techniques
What will you get from this session?
1. Insights into SAP testing best practices
2. Heatmap utilization for testing
3. Optimization of testing processes
4. Demo
Topics covered:
Execution from the test manager
Orchestrator execution result
Defect reporting
SAP heatmap example with demo
Speaker:
Deepak Rai, Automation Practice Lead, Boundaryless Group and UiPath MVP
UiPath Test Automation using UiPath Test Suite series, part 3DianaGray10
Welcome to UiPath Test Automation using UiPath Test Suite series part 3. In this session, we will cover desktop automation along with UI automation.
Topics covered:
UI automation Introduction,
UI automation Sample
Desktop automation flow
Pradeep Chinnala, Senior Consultant Automation Developer @WonderBotz and UiPath MVP
Deepak Rai, Automation Practice Lead, Boundaryless Group and UiPath MVP
2. Contents
Definition
Meaning of PCR.
Purpose of PCR
Components of PCR.
Steps of PCR
Advantages
Disadvantages
REAL TIME PCR
Definition
Detection methods in real time PCR
Applications.
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3. Definition :
Polymerase chain reaction is a technique used in molecular
biology to amplify a single copy or a few copies of a segment of
DNA ,generating thousands to millions of copies of a particular
DNA sequence.
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4. What is meaning of PCR ?
P stands for polymerase
because the only enzyme
used in this reaction is DNA
polymerase
R stands for reaction
because the reaction
takes place in process.
C stands for chain because
the products of the 1st
reaction become
substrates of the following
one, and so on
PCR
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5. Purpose:
To amplify a lot of double stranded DNA molecules with same size
and sequence by enzymatic method and cycling condition.
Developed in 1983 by Karry Mullis. In 1993 Mullis was awarded the
Nobel prize in chemistry for his work on PCR.
PCR is now considered as a basic tool for the molecular biologist.
As is a photocopier a basic requirement in an office, so is the PCR
machine in a molecular biology laboratory!
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6. Components of PCR
DNA template: It is DNA segment to be amplified.
Two primers: Primers are synthetic DNA strands of about 18 to 25
nucleotides complimentary to 3’ end of template strand.
Forward primer: It is complimentary to the 3’ end of antisense
strand (3’-5’).
Reverse primer: It is complimentary to the 3’ end of sense strand
(5’-3’).
Taq polymerase: Taq polymerase adds nucleotides complimentary to
template strand and synthesis new strand of DNA. It is isolated from
Thermus aquaticus( heat resistant bacteria). It is used because of
high temperature stability.
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7. Nucleotides(dNTPs): All types of nucleotides are “building
blocks’’ for new DNA strands and essential for reaction. It
includes Adenine(A), Guanine(G), Thymine(T), Cytosine(C).
Buffer solution: providing a suitable chemical environment for
optimum activity and stability of the DNA polymerase.
Divalent cations: Polymerases require free divalent cations
usually Mg+2 for activity. It act as cofactor in the catalytic
addition of dNTPs.
Monovalent ions(K+): It promotes primer annealing.
PCR Machine: a thermal cycler.
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9. Steps of PCR
There are three major steps in a PCR, which are repeated for 30
or 40 cycles.
This is done on an automated cycler ,which can heat and cool the
tubes with the reaction mixture in a very short time.
The three major steps are as follows:
1) Denaturation at 94oC
2) Annealing at 540C
3) Extension at 72oC
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10. Denaturation at 94oC
During the these step, the reaction mixture is heated to 94OC for
1 min, which causes separation of DNA double stranded. Now
each strand acts as template for synthesis of complimentary
strand.
The Hydrogen bonds between the two strands breaks down and
the two strands separates.
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11. Annealing at 54oC
This step consist of cooling of reaction mixture after denaturation
step to 54oC, which causes annealing of primers to separated strand of
DNA.
The length and GC-content of the primer should be sufficient for
stable binding with template .Guanine pairs with cytosine with three
hydrogen bonds. Thus, higher GC content results in stronger binding.
Time taken to anneal is 45 second.
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12. Extension at 72oC
Taq polymerase binds to the template DNA and starts adding
nucleotides that are complementary to the first strand.
This happens at 72oC as it is
the optimum temperature
for Taq polymerase.
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13. Advantages :
Rapid and easy to perform.
Small amount of DNA is required per test.
Result obtained more quickly.
Usually not necessary to use radioactive material for PCR.
PCR can be used to detect point mutations
Making it possible to amplify DNA from degraded samples.
It is very accurate ,especially for determining various
diseases, leading to better diagnoses.
It is most specific ,sensitive.
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14. Disadvantages :
Target DNA sequence must be known.
Errors made by polymerase.
High degree of operator skill required.
High equipment cost.
High sterile environment should be provided.
Chances of contamination.
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15. What is the need of real time PCR ?
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16. Real Time PCR
Definition:
A real time polymerase chain reaction is a laboratory technique of
molecular biology based on the polymerase chain reaction. It monitors
the amplification of a targeted DNA molecule during the PCR, i.e. in real
time, and not at its end, as in conventional PCR.
It is also called as quantitative real time polymerase chain reaction.
PRINCIPLE: The amount of the nucleic acid present into the sample is
quantified using the fluorescent dye or using the fluorescent labeled
oligos.
Real Time PCR is sensitive and reliable method for detection and
quantification of nucleic acid levels.
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17. Detection methods in Real Time PCR:
(1) By using sequence specific fluorescent probes
(2) By using non specific fluorescent dyes
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18. 1) By using sequence specific fluorescent probes
Fluorescent reporter probes detect only the DNA containing the probe
sequence ; therefore ,use of the reporter probe significantly increases
specificity.
It is hydrolysis probe which bear a reporter dye, often fluorescein at its 5’
end and a quencher attached to the 3’ end of the oligonucleotide.
Under normal condition, the probe remain coiled on itself bringing the
fluorescence dye near the quencher, which inhibits or quenches of
fluorescent single of the dye so it is also known as FRET ( fluorescence
resonance energy transfer).
During the annealing stage of the rtPCR both probe and primers anneal to
separated strand of DNA.
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19. • As the taq-polymerase start to synthesize new DNA strand in the
extension stage, and once the polymerase reaches the probe ,its 5’-
3’exonuclease degrades the probe, physically separating the fluorescent
reporter from the quencher, resulting in an increase in fluorescence.
• The increase in PCR product is proportional to amount of fluorescence.
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21. 2) By using non specific fluorescent dyes
Non specific fluorescent dye binds to all double stranded DNA product
in real time PCR, causing fluorescence of the dye.
An increase in DNA product during PCR therefore leads to an increase in
fluorescence intensity and is measured at each cycle, thus allowing DNA
concentrations to be quantified.
SYBR Green 1 ,SYBR Green 2, EVA green, LC green dyes are used in these
process.
SYBR Green is most widely used for its higher signal intensity.
The reaction is prepared as usual, with the addition of fluorescent dsDNA
dye.
The reaction is run in a real time PCR instrument, and after each the levels
of fluorescence are measured with a detector. 21
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23. Detection :
The detection is based on fluorescence technology.
The specimen is first kept in proper well and subjected to thermal cycle
like in normal PCR.
The machine, however ,in the real time PCR is subjected to tungsten or
halogen source that lead to fluoresce the marker added to the sample and
the signal is amplified with the amplification of copy number of sample
DNA.
The emitted signal is detected by an detector and sent to computer after
conversion into digital signal that is displayed on screen.
The signal can be detected when it comes up the threshold level.
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24. ct value: It is defined
as number of cycles
required for the
fluorescent signal to
cross the threshold.
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25. Applications :
There are different applications for real time PCR as follows:
The use of PCR in clinical settings can be broadly divided into three
categories:
1) To amplify human genes to check for mutations.
2) To amplify microbial genes in a sample.
3) To amplify human gene from a limited sample for creating a
complete DNA profile of an individual.
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26. Diagnosis of Virus
The virus contain mRNA as a genetic material so that for detection of
virus is done by coupling of real time PCR with a procedure called
reverse transcription.
In this method , RNA is first
transcribed into complementary
DNA(cDNA) by reverse transcriptase
from total mRNA. The cDNA is then
used as the template for the real
time PCR reaction.
It is also used in diagnosis of genetic
disease, cancer etc.
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27. Gene Expression
It is most sensitive method for the detection and quantification of gene
expression levels.
It is used for determining how the gene expression and gene changes
over time ,such as in:
1) The response of tissue and cell cultures to an administration of a
pharmacological agent
2) response to change in environmental conditions.
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28. Agriculture
Detection of phytopathogens:
The agricultural industry is constantly striving to produce plant
propagules that are free of pathogens in order to prevent economic
losses and safeguard health.
Discrimination between the DNA of the pathogen and the plant is based
on the amplification of specific sequences in ribosomal RNA gene’s
coding area by real time PCR.
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29. Genetically modified organisms (GMO)
A genetically modified organism(GMO) is a living organism e.g. a plant
,whose genetic composition has been altered by means of gene technology.
The genetic modification usually involves insertion of a piece of DNA , into
the genome of the organism to be modified.
These smaller piece of DNA is usually taken from other naturally occurring
organisms.
Specific primers are that amplify the gene sequences used during the
process of engineering the vector.
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30. In Gene therapy:
In gene therapy as a drug delivery system there are two important
parameter have to be analyzed :
1) The expression level of the therapeutic gene and
2) The distribution of the drug in different organs.
For the control of both parameters , real time PCR is used.
Microbiological uses:
used by microbiologists in the fields of food safety, food spoilage and
fermentation and for the microbial risk assessment of water quality.
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