Polymerase chain reaction (PCR) is a method widely used 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. PCR was invented in 1984 by the American biochemist Kary Mullis at Cetus Corporation. It is fundamental to much of genetic testing including analysis of ancient samples of DNA and identification of infectious agents. Using PCR, copies of very small amounts of DNA sequences are exponentially amplified in a series of cycles of temperature changes. PCR is now a common and often indispensable technique used in medical laboratory and clinical laboratory research for a broad variety of applications including biomedical research and criminal forensics

1. Polymerase
Chain Reaction

2. 2
PCR
● PCR – 37th Anniversary in 2020
● Now, a ubiquitous laboratory tool in all laboratories
● Limited use outside laboratories
● Efforts to make the system
○ Portable (size reduction)
○ Affordable
○ Energy efficient / Electricity-free
miniPCRTM
Palm PCRTM

3. PCR has amplified itself in multiple fields!!
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International
Space Station
Infectious
Diseases
Inherited Genetic
Diseases
Neonatal Diseases
& Sex
Determination
Cancer diagnosis,
prognosis,
monitoring
Tracking Animal
Migration
Food Pathogen
and GM
verification
Organ
Transplantation
Archival Material

4. 4
PCR in
Genomic
Research DNA
Fingerprinting
Genotyping Gene Expression Cell line
Validation
PCR Cloning DNA Methylation
Assay
DNA Sequencing Library
Preparation

5. PCR
Roche LightCycler (real-time PCR)

6. It is NOT that simple.
Even with the simplest PCR reaction, things can go wrong! 6

7. Important Considerations
If assay design is robust, you can
interpret your results with confidence.
E.g. Use of positive control, negative
control, endogenous and exogenous
internal controls
Interpretation of PCR Results
D
Visualization / Detection /
Analysis of PCR products
Post-PCR Analysis
C
Depends on the purpose /
goal of the Assay
PCR Assay Design
A
Based on Assay Design;
Troubleshooting
Performing the PCR Test
B

8. 8

9. What can go wrong?
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No Amplification of Target
Weak Amplification of Target
Non-Specific Band Amplification

10. 1
Wrong
Interpretation
E.g. Covid-19 RT-PCR Results
False Negative Results
 Improper sample collection/transport
 Insufficient amount of specimen
 Degradation of nucleic acids (typically RNA) during shipping or storage
 Specimen collected prior to onset of symptoms or late in illness
 Quantity of organisms is below detection limit
 Non-homogeneous distribution of the organism of interest
 Presence of amplification inhibitors in the specimen
 Laboratory processing/testing errors

11. 1
Wrong
Interpretation
E.g. Covid-19 RT-PCR Results
False Positive Results
 Detecting contaminants introduced during specimen collection,
transport or processing
 Detecting organisms representative of normal flora near specimen
collection site and contaminants in lab
 Mislabeling
 Specimen mix-up

12. PCR - A Perception of the Molecular Approach
BioTecNika’s Course on PCR
The Need of the hourThe nitty-gritties to be
considered while
performing PCR
Seemingly simple, yet
complex in execution
The importance of
PCR technique
Highly versatile and
Fast-evolving technique
‘PCR’ has no
competition

13. PCR Laboratory Practices - Protocols and Precaution
PCR - A Perception of the Molecular Approach
1
2
3
4
Basics of PCR - Introduction, Principle, Steps
Types of PCR and their specific applications
Required components of PCR and chemistry - PCR template, primer design
Sample preparation, quality control and optimization of PCR parameters
5

14. PCR Multiplexing
PCR - A Perception of the Molecular Approach
6
7
8
9
Detection of PCR products and analysis of results
PCR applications: PCR-based genome analysis
Gene expression studies and Micro RNA assays
Real-time PCR - Fundamentals, Assay design, Normalization
10

15. Demo on Performing a PCR
PCR - A Perception of the Molecular Approach
Quantitative PCR and Bioinformatics
PCR in Diagnostic testing
PCR in COVID-19 Detection
Anomalies of PCR as a Research Tool-Case studies
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12
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16. Genomic/Mol-bio based Research
PCR an integral part
The PCR Industry itself!
Designing and Developing PCR systems
Various Sectors that use PCR
Medical, Diagnostic, Forensic, Food and Beverage Industries, Agriculture,
Dairy and Poultry, Toxicologists, Environmentalists,
Ecologists/Conservationists, Climate change scientists, Archaeologists
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How will it help you?

17. “Polymerase Chain Reaction Market to Exhibit 8.3% CAGR by
2026: Advent of Digital PCR is Promoting Growth
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- Fortune Business Insights™ (June 08, 2020)

18. • Infographic Style
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