The polymerase chain reaction (PCR) is a technique for amplifying DNA segments. It involves denaturing DNA, annealing primers to the single strands, and extending the primers to replicate the DNA segment. Kary Mullis developed PCR in 1983 and won the Nobel Prize for it. PCR consists of cycles of heating and cooling DNA which allows exponential amplification of specific DNA regions defined by primer binding sites. It has many applications in research, forensics, medicine and more.
PCR is a biochemical technology in molecular biology. This technique used in DNA cloning for sequencing, gene cloning and manipulating, diagnosis of hereditary diseases.
What is PCR?
History of PCR
Components of PCR
Principles of PCR
Basic Requirements
Instrumentation
PCR Programme
Advantages of PCR
Applications of PCR
Conclusion
References
PCR is a biochemical technology in molecular biology. This technique used in DNA cloning for sequencing, gene cloning and manipulating, diagnosis of hereditary diseases.
What is PCR?
History of PCR
Components of PCR
Principles of PCR
Basic Requirements
Instrumentation
PCR Programme
Advantages of PCR
Applications of PCR
Conclusion
References
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.
The major agent of causes meningitis are :
1- 9802 gene fragment in Streptococcus pneumoniae.
2- omp P6 gene in Haemophilus influenzae .
3- ctrA gene in Neisseria meningitidis .
We can not detect these agents by Conventional lab. methods , and therefore we use PCR Techniques to ensure detect these agents
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.
Polymerase chain reaction .... Very important topic which is explained in very simple and convenient way. Learning objectives and references are given in the presentation for the detailed learning. The presentation was Guided by Dr Shilpa Jain and made by Ms. Nidhi Argade.
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.
PCR explained in simple terms - A T G & C of PCR - Question and answers PCRajithnandanam
www.technologyinscience.blogspot.com
PCR - polymerase chain reaction explained in simple question answer format. Type in your doubts on PCR in the comment.
video on YouTube
https://www.youtube.com/watch?v=NLmonT3GUHE
https://www.youtube.com/watch?v=NLmonT3GUHE
Polymerase Chain Reaction, PCR
is a method widely used in molecular biology to rapidly make millions to billions of copies of a specific DNA sample allowing scientists to take a very small sample of DNA and amplify it to a large enough amount to study in detail.
Polymerase chain reaction (abbreviated PCR) is a laboratory technique for rapidly producing (amplifying) millions to billions of copies of a specific segment of DNA, which can then be studied in greater detail. PCR involves using short synthetic DNA fragments called primers to select a segment of the genome to be amplified, and then multiple rounds of DNA synthesis to amplify that segment. This slides introduces pcr importances ,uses and steps of pcr.
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.
The major agent of causes meningitis are :
1- 9802 gene fragment in Streptococcus pneumoniae.
2- omp P6 gene in Haemophilus influenzae .
3- ctrA gene in Neisseria meningitidis .
We can not detect these agents by Conventional lab. methods , and therefore we use PCR Techniques to ensure detect these agents
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.
Polymerase chain reaction .... Very important topic which is explained in very simple and convenient way. Learning objectives and references are given in the presentation for the detailed learning. The presentation was Guided by Dr Shilpa Jain and made by Ms. Nidhi Argade.
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.
PCR explained in simple terms - A T G & C of PCR - Question and answers PCRajithnandanam
www.technologyinscience.blogspot.com
PCR - polymerase chain reaction explained in simple question answer format. Type in your doubts on PCR in the comment.
video on YouTube
https://www.youtube.com/watch?v=NLmonT3GUHE
https://www.youtube.com/watch?v=NLmonT3GUHE
Polymerase Chain Reaction, PCR
is a method widely used in molecular biology to rapidly make millions to billions of copies of a specific DNA sample allowing scientists to take a very small sample of DNA and amplify it to a large enough amount to study in detail.
Polymerase chain reaction (abbreviated PCR) is a laboratory technique for rapidly producing (amplifying) millions to billions of copies of a specific segment of DNA, which can then be studied in greater detail. PCR involves using short synthetic DNA fragments called primers to select a segment of the genome to be amplified, and then multiple rounds of DNA synthesis to amplify that segment. This slides introduces pcr importances ,uses and steps of pcr.
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
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
MOLECULAR TOOLS IN DIAGNOSIS AND CHARACTERIZATION OF INFECTIOUS DISEASES tawheedshafi
The future of the molecular diagnostics of infectious diseases will undoubtedly be focused on a marked increase in the amount of information detected with remarkably simplified, rapid platforms that will need complex software analysis to resolve the data for use in clinical decision-making.
MATATAG CURRICULUM: ASSESSING THE READINESS OF ELEM. PUBLIC SCHOOL TEACHERS I...NelTorrente
In this research, it concludes that while the readiness of teachers in Caloocan City to implement the MATATAG Curriculum is generally positive, targeted efforts in professional development, resource distribution, support networks, and comprehensive preparation can address the existing gaps and ensure successful curriculum implementation.
Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
Unit 8 - Information and Communication Technology (Paper I).pdfThiyagu K
This slides describes the basic concepts of ICT, basics of Email, Emerging Technology and Digital Initiatives in Education. This presentations aligns with the UGC Paper I syllabus.
Safalta Digital marketing institute in Noida, provide complete applications that encompass a huge range of virtual advertising and marketing additives, which includes search engine optimization, virtual communication advertising, pay-per-click on marketing, content material advertising, internet analytics, and greater. These university courses are designed for students who possess a comprehensive understanding of virtual marketing strategies and attributes.Safalta Digital Marketing Institute in Noida is a first choice for young individuals or students who are looking to start their careers in the field of digital advertising. The institute gives specialized courses designed and certification.
for beginners, providing thorough training in areas such as SEO, digital communication marketing, and PPC training in Noida. After finishing the program, students receive the certifications recognised by top different universitie, setting a strong foundation for a successful career in digital marketing.
The simplified electron and muon model, Oscillating Spacetime: The Foundation...RitikBhardwaj56
Discover the Simplified Electron and Muon Model: A New Wave-Based Approach to Understanding Particles delves into a groundbreaking theory that presents electrons and muons as rotating soliton waves within oscillating spacetime. Geared towards students, researchers, and science buffs, this book breaks down complex ideas into simple explanations. It covers topics such as electron waves, temporal dynamics, and the implications of this model on particle physics. With clear illustrations and easy-to-follow explanations, readers will gain a new outlook on the universe's fundamental nature.
A review of the growth of the Israel Genealogy Research Association Database Collection for the last 12 months. Our collection is now passed the 3 million mark and still growing. See which archives have contributed the most. See the different types of records we have, and which years have had records added. You can also see what we have for the future.
A workshop hosted by the South African Journal of Science aimed at postgraduate students and early career researchers with little or no experience in writing and publishing journal articles.
This slide is special for master students (MIBS & MIFB) in UUM. Also useful for readers who are interested in the topic of contemporary Islamic banking.
Delivering Micro-Credentials in Technical and Vocational Education and TrainingAG2 Design
Explore how micro-credentials are transforming Technical and Vocational Education and Training (TVET) with this comprehensive slide deck. Discover what micro-credentials are, their importance in TVET, the advantages they offer, and the insights from industry experts. Additionally, learn about the top software applications available for creating and managing micro-credentials. This presentation also includes valuable resources and a discussion on the future of these specialised certifications.
For more detailed information on delivering micro-credentials in TVET, visit this https://tvettrainer.com/delivering-micro-credentials-in-tvet/
June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
2. What is PCR?
The polymerase chain reaction (PCR)
is a rapid, inexpensive, popular
molecular biology technique for
enzymatically replicating DNA
(without using a living organism such
as E.coli or yeast) & produces
microgram amounts of DNA from
minute quantities of template. This
technique allows a small amount of
the DNA molecule to be amplified
many times in an exponential
manner.
3. History
PCR technique was developed in
1983 by Kary Mullis. In 1993,
Mullis was awarded the Nobel
prize in Chemistry along with
Michael Smith for his work on
PCR.
Dr. Kary Mullis stated it “lets
you pick the piece of DNA you
are interested in & have as
much of it as you want”.
4. Steps
The PCR process consists of a series
of twenty to thirty-five cycles. Each
consists of three steps:
1. Denaturation of dsDNA
2. Annealing of the primers
3. Extension of the primers
5. The double-stranded DNA has to be heated
to 94-96 °C in order to separate the
strands. This is called denaturing; it breaks
apart the hydrogen bonds that connect the
two DNA strands. Prior to the first cycle,
the DNA is often denatured for an extended
time to ensure that both the template DNA
& the primers have completely separated &
are now single-strand only.
Time 1-2 minutes up to 5 minutes.
Also Taq–polymerase is activated by this
step.
Denaturing
6. After separating the DNA strands, the
temperature is lowered so the primers
can attach themselves to the single
DNA strands. This step is called
annealing.
The temperature of this stage depends
on the primers & usually 50 °C below
their melting temperature (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.
Annealing
7. Extension
Finally, the DNA-polymerase has to fill in
the missing strands. It starts at the
annealed primer & works its way along
the DNA strand. This step is called
extension. The extension temperature
depends on the DNA-polymerase.
The time for this step depends both on the
DNA-polymerase itself & on the length of
the DNA fragment to be amplified.
As a rule-of-thumb, 1minute per 1 kbp.
8. Stages
The PCR process can be divided into three
stages:
1. Exponential amplification: At every cycle,
the amount of product is doubled
(assuming 100% reaction efficiency). This
reaction is very sensitive, only minute
quantities of DNA need to be present.
2. Leveling off stage: The reaction slows as the
DNA polymerase loses activity & as
consumption of reagents such as dNTPs &
primers causes them to become limiting.
3. Plateau: No more product accumulates due
to exhaustion of reagents & enzymes.
9. Components
• DNA template or cDNA which contains the region
of the DNA fragment to be amplified
• Two primers which determine the beginning & end
of the region to be amplified
• Taq polymerase (DNA polymerase) extends primers
& copies the region to be amplified
• Nucleotides from which the DNA-polymerase for
new DNA
• Buffers provides a suitable chemical environment for
the DNA-polymerase
• Mg²+ is required as co-factor for the thermostable
DNA polymerase
• Gelatin & Triton X-100 stabilise the DNA
polymerase
• dNTPs provide initial excess required for
incorporation into DNA
10. PRIMERS
These are short, artificial DNA strands not
more than fifty, usually 18-25 bp
nucleotides that are complementary to the
beginning & end of the DNA fragment to be
amplified.
Both primers which used in the reaction,
should have similar annealing temperatures
with a minimal degree of
self-complementarity in order to avoid the
formation of secondary structures & no
complementarity to each other so that the
primers dimers will not formed.
11. The PCR reaction is
carried out in a
machine that heats &
cools the reaction tubes
within it, to the precise
temperature required
for each step of the
reaction, is called
Thermal Cycler.
12. The PCR product
can be identified
by its size using
agarose gel
electrophoresis.
13. Applications
✓ Diagnosis & screening of genetic diseases
& cancer
✓ Rapid detection of slowly growing
microorganisms & viruses
✓ HLA typing in transplantation
✓Analysis of DNA in archival material
✓ DNA fingerprinting in forensic science
✓ Preparation of nucleic acid probes
✓ Clone screening, mapping & sub-cloning
✓ Paternity test
14. Disadvantages
▪ requires costly instruments
▪ adequate space with aircondition,
dehumidifier, laminar flow facilities
▪ costly & not all people can afford
to do test
▪ requires trained, experienced,
qualified manpower & technologists
▪ false positive & false negative
results may lower specificity &
sensitivity
▪ limited scope for diagnosis of
diseases
15. Practical Modification
These are some practical modifications
to PCR technique:
• Nested PCR
• Inverse PCR
• Reverse Transcriptase RT-PCR
• Asymetric PCR
• Quantitative PCR
• Real time PCR
• Touchdown PCR
• Colony PCR
• Allele-specific PCR