The document discusses the principle and applications of polymerase chain reaction (PCR). It begins by explaining how PCR is used to amplify DNA sequences and was discovered by Kary Mullis. It then describes the three main steps of PCR (denaturation, annealing, elongation) and how they are used to exponentially increase copies of DNA. Applications discussed include medicine, infectious disease detection, genetic testing, plant science, veterinary science, 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
Diagnostic polymerase chain reaction (PCR) is an extremely powerful, rapid method for diagnosis of microbial infections and genetic diseases, as well as for detecting microorganisms in environmental and food samples.
However, the usefulness of diagnostic PCR is limited, in part, by the presence of inhibitory substances in complex biological samples, which reduce or even block the amplification capacity of PCR in comparison with pure solutions of nucleic acids .
In general, diagnostic PCR may be divided into four steps: (1) sampling, (2) sample preparation, (3) nucleic acid amplification, and (4) detection of PCR products
Diagnostic polymerase chain reaction (PCR) is an extremely powerful, rapid method for diagnosis of microbial infections and genetic diseases, as well as for detecting microorganisms in environmental and food samples.
However, the usefulness of diagnostic PCR is limited, in part, by the presence of inhibitory substances in complex biological samples, which reduce or even block the amplification capacity of PCR in comparison with pure solutions of nucleic acids .
In general, diagnostic PCR may be divided into four steps: (1) sampling, (2) sample preparation, (3) nucleic acid amplification, and (4) detection of PCR products
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
Diagnostic polymerase chain reaction (PCR) is an extremely powerful, rapid method for diagnosis of microbial infections and genetic diseases, as well as for detecting microorganisms in environmental and food samples.
However, the usefulness of diagnostic PCR is limited, in part, by the presence of inhibitory substances in complex biological samples, which reduce or even block the amplification capacity of PCR in comparison with pure solutions of nucleic acids .
In general, diagnostic PCR may be divided into four steps: (1) sampling, (2) sample preparation, (3) nucleic acid amplification, and (4) detection of PCR products
Diagnostic polymerase chain reaction (PCR) is an extremely powerful, rapid method for diagnosis of microbial infections and genetic diseases, as well as for detecting microorganisms in environmental and food samples.
However, the usefulness of diagnostic PCR is limited, in part, by the presence of inhibitory substances in complex biological samples, which reduce or even block the amplification capacity of PCR in comparison with pure solutions of nucleic acids .
In general, diagnostic PCR may be divided into four steps: (1) sampling, (2) sample preparation, (3) nucleic acid amplification, and (4) detection of PCR products
PCR is a revolutionary molecular biology technique used for enzymatically replicating DNA . This technique allows a small amount of DNA molecule to be amplified many times in an exponential manner . It is commonly used in medical and biological research labs for variety of tasks such as detection of hereditary disease , identification of genetic fingerprints diagnosis of infectious disease , cloning of genes and paternity testing .
Each reaction cycle doubles the amount of DNA – a standard PCR sequence of 30 cycles creates over 1 billion copies . The thermostability of DNA polymerases is defined by how long they remain active at the extreme range of temperatures used in PCR.
There have been various thermostable polymerases identified to date, each with its optimal temperature for activity and a unique half-life profile at temperatures greater than 95°C. For example, the half-life of Taq polymerase at 95°C is 40 minutes, whereas the half-life of the hyperthermophilic Deep Vent DNA polymerase extracted from the Pyrococcus species GB-D is several hours at 98–100°C. Polymerase processivity is defined as the number of consecutive nucleotides a single enzyme can incorporate before being dislodged from the DNA template.
At 75°C, native Taq polymerases can typically amplify DNA at a rate of 10–45 nucleotides per second - that’s approximately 2 kilobases per minute!
Some DNA polymerases have been engineered to improve their binding domain, thus making them more stable than conventional Taq. For example, KAPA2G polymerase has a speed of ~150 nucleotides per second - 3-fold higher than Taq. Direct PCR cloning methods include TA and GC cloning, as well as TOPO® Cloning, and enable direct cloning of PCR fragments. For example, the TA cloning approach takes advantage of the 3’ A overhang naturally added to products by Taq polymerase following PCR. The resulting sticky ends then enable recombination with DNA fragments containing 3’ T overhangs, such as linearized vectors.
During indirect PCR cloning, the PCR products are modified prior to recombination with other DNA sequences. For example, in restriction cloning, restriction sites are frequently introduced via PCR to enable restriction digestion and ligation with linearized vectors. PCR mutagenesis is a technique used to generate site-directed sequence changes such as base substitutions, inserts and deletions.
To insert a single point mutation via mutagenesis, for example, PCR primers are designed that contain the desired base change, usually in the middle of the primer sequence. PCR is then performed with the mutagenic primers and a high-fidelity DNA polymerase, which results in the incorporation of the desired mutation into the original sequence.Allele-specific PCR is used to detect sequence variations and ultimately determine the genotype of an organism.
For allele-specific PCR, primers are designed to flank the region of interest. The most common application of PCR is gene expression analysis
SLIDE CONTAIN BREIF NOTE ON PCR. IT CONTAINS 21 SLIDES INCLUDING, WHAT IS PCR? COMPONENTS, WORKING MECHANISM, APPLICATIONS, CONCLUSION, AND SOME REFRENCES, HISTORY ALSO
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
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.
PCR is a revolutionary molecular biology technique used for enzymatically replicating DNA . This technique allows a small amount of DNA molecule to be amplified many times in an exponential manner . It is commonly used in medical and biological research labs for variety of tasks such as detection of hereditary disease , identification of genetic fingerprints diagnosis of infectious disease , cloning of genes and paternity testing .
Each reaction cycle doubles the amount of DNA – a standard PCR sequence of 30 cycles creates over 1 billion copies . The thermostability of DNA polymerases is defined by how long they remain active at the extreme range of temperatures used in PCR.
There have been various thermostable polymerases identified to date, each with its optimal temperature for activity and a unique half-life profile at temperatures greater than 95°C. For example, the half-life of Taq polymerase at 95°C is 40 minutes, whereas the half-life of the hyperthermophilic Deep Vent DNA polymerase extracted from the Pyrococcus species GB-D is several hours at 98–100°C. Polymerase processivity is defined as the number of consecutive nucleotides a single enzyme can incorporate before being dislodged from the DNA template.
At 75°C, native Taq polymerases can typically amplify DNA at a rate of 10–45 nucleotides per second - that’s approximately 2 kilobases per minute!
Some DNA polymerases have been engineered to improve their binding domain, thus making them more stable than conventional Taq. For example, KAPA2G polymerase has a speed of ~150 nucleotides per second - 3-fold higher than Taq. Direct PCR cloning methods include TA and GC cloning, as well as TOPO® Cloning, and enable direct cloning of PCR fragments. For example, the TA cloning approach takes advantage of the 3’ A overhang naturally added to products by Taq polymerase following PCR. The resulting sticky ends then enable recombination with DNA fragments containing 3’ T overhangs, such as linearized vectors.
During indirect PCR cloning, the PCR products are modified prior to recombination with other DNA sequences. For example, in restriction cloning, restriction sites are frequently introduced via PCR to enable restriction digestion and ligation with linearized vectors. PCR mutagenesis is a technique used to generate site-directed sequence changes such as base substitutions, inserts and deletions.
To insert a single point mutation via mutagenesis, for example, PCR primers are designed that contain the desired base change, usually in the middle of the primer sequence. PCR is then performed with the mutagenic primers and a high-fidelity DNA polymerase, which results in the incorporation of the desired mutation into the original sequence.Allele-specific PCR is used to detect sequence variations and ultimately determine the genotype of an organism.
For allele-specific PCR, primers are designed to flank the region of interest. The most common application of PCR is gene expression analysis
SLIDE CONTAIN BREIF NOTE ON PCR. IT CONTAINS 21 SLIDES INCLUDING, WHAT IS PCR? COMPONENTS, WORKING MECHANISM, APPLICATIONS, CONCLUSION, AND SOME REFRENCES, HISTORY ALSO
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
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.
Similar to PCR its principle and application related to botany.pptx (20)
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
The increased availability of biomedical data, particularly in the public domain, offers the opportunity to better understand human health and to develop effective therapeutics for a wide range of unmet medical needs. However, data scientists remain stymied by the fact that data remain hard to find and to productively reuse because data and their metadata i) are wholly inaccessible, ii) are in non-standard or incompatible representations, iii) do not conform to community standards, and iv) have unclear or highly restricted terms and conditions that preclude legitimate reuse. These limitations require a rethink on data can be made machine and AI-ready - the key motivation behind the FAIR Guiding Principles. Concurrently, while recent efforts have explored the use of deep learning to fuse disparate data into predictive models for a wide range of biomedical applications, these models often fail even when the correct answer is already known, and fail to explain individual predictions in terms that data scientists can appreciate. These limitations suggest that new methods to produce practical artificial intelligence are still needed.
In this talk, I will discuss our work in (1) building an integrative knowledge infrastructure to prepare FAIR and "AI-ready" data and services along with (2) neurosymbolic AI methods to improve the quality of predictions and to generate plausible explanations. Attention is given to standards, platforms, and methods to wrangle knowledge into simple, but effective semantic and latent representations, and to make these available into standards-compliant and discoverable interfaces that can be used in model building, validation, and explanation. Our work, and those of others in the field, creates a baseline for building trustworthy and easy to deploy AI models in biomedicine.
Bio
Dr. Michel Dumontier is the Distinguished Professor of Data Science at Maastricht University, founder and executive director of the Institute of Data Science, and co-founder of the FAIR (Findable, Accessible, Interoperable and Reusable) data principles. His research explores socio-technological approaches for responsible discovery science, which includes collaborative multi-modal knowledge graphs, privacy-preserving distributed data mining, and AI methods for drug discovery and personalized medicine. His work is supported through the Dutch National Research Agenda, the Netherlands Organisation for Scientific Research, Horizon Europe, the European Open Science Cloud, the US National Institutes of Health, and a Marie-Curie Innovative Training Network. He is the editor-in-chief for the journal Data Science and is internationally recognized for his contributions in bioinformatics, biomedical informatics, and semantic technologies including ontologies and linked data.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
This pdf is about the Schizophrenia.
For more details visit on YouTube; @SELF-EXPLANATORY;
https://www.youtube.com/channel/UCAiarMZDNhe1A3Rnpr_WkzA/videos
Thanks...!
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
2. Polymerase chain reaction
Polymerase chain reaction (PCR) is used to
amplify a DNA sequence to produce millions of
copies.
Kary Mullis discovered the PCR and got Nobel
Prize in Chemistry in 1983 for his discovery.
Since then, PCR has been used in various
applications in medicine, animal science, plant
science, food science etc.
3. Principle of the technique
Whole process of PCR involves three main events, Denaturation, Annealing
and Elongation. A DNA fragment of interest is used as a template and a pair of
primers which are short oligonucleotides complimentary to the both strands of
the template DNA. The purpose of primer is to initiate the DNA synthesis in
the direction of 5‟ to 3‟.
The number of amplified DNA or the amplicons increases exponentially per
cycle thus one molecule of DNA gives rise to 2,4,8,16 and so forth. This
continuous doubling is carried out by a specific enzyme called DNA
polymerase which sits at the unfinished double stranded DNA created by
template DNA and primer.
For further extension of the DNA, the polymerase enzyme require supply of
other DNA-building blocks such as the nucleotides consisting of four bases
Adenine (A), Thymine (T), Cytosine (C) and Guanine (G). The template,
primer, polymerase and four bases are the main components for polymerase
chain reaction.
4.
5. Methodology:
PCR has three major events (Denaturation, Annealing and Elongation) to
complete the amplification process (Figure 15.1). The complete process of
PCR is as follows-
Initial denaturation: Heating the PCR mixture at 94°C to 96°C for 10min to
ensure complete denaturation of template DNA. It is followed by the cyclic
events which has different steps as described below:
Denaturation: This is the first step in which the double stranded DNA
template is denatured to form two single strand by heating at 95°C for 15-30
secs.
Annealing: This is the annealing step where at lower temperature (usually 50-
650C) primers are allowed to bind to template DNA, annealing time is 15-30
secs and it depends on the length and bases of the primers. Generally annealing
temperature is about 3-5°C below the melting temperature (Tm) of the pair of
the primers is used. PCR has three major events (Denaturation, Annealing and
Elongation) to complete the amplification process (Figure 15.1). The complete
process of PCR is as follows-
6. Methodology:
Elongation: This is the synthesis step where the polymerase
perform synthesis of new strand in the 5‟ to 3‟ direction using
primer and deoxyribonucleoside triphosphates (dNTPs). An
average DNA polymerase adds about 1,000 bp/minute. Step 1,2,3
makes one cycle and in general 35-40 such cycles are performed
in a typical PCR amplification.
After the cycles are completed, the reaction is held at 70-74°C for
several minutes to allow final extension of the remaining DNA to
be fully extended.
Termination: The reaction is complete and the resulting
amplified nucleic acids are held at a low temperature (~4°C) until
analysis.
7. Instrumentation:
Thermal cycler is the instrument that carries out the
amplification via polymerase chain reaction (Figure
15.2). Usually the three main events are repeated for 30-
40 cycles to obtain detectable amount of product at the
end of the cycles.
The automated system performs the cyclic temperature
changes required for enzymatic amplification of specific
DNA segments in vitro using this PCR.
The device has a thermal block with holes where tubes
containing reaction mixtures can be inserted. The cycler
varies the temperature of the block in discrete, pre-
programmed steps using peltier effect
8. . Primers:
A primer is a short oligonucleotide that serves as a starting point for DNA
synthesis. In PCR, two primers are required to bind to each of the single
stranded DNA (obtained after denaturation) flanking the target sequence. These
are called Forward and Reverse primers. They primers are designed in such a
way that they have a sequence complimentary to the sequence in the template
DNA. Two restriction enzymes sites are added at the 5‟ end of each of the
primer to facilitate cloning. The chosen restriction enzymes will not cut DNA
fragment (non-cutters). Typically 3 to 4 nucleotides are added at the end of the
restriction sites to allow efficient cutting by restriction enzymes.
9. Primer Designing and criteria:
For a specific amplifications in PCR, good primer design is
essential. The following parameters needs to be considered while
designing a primer:
1. Primer length: Oligonucleotides between 18-24 bases is the
ideal length which is long enough for adequate specificity and
short enough for primers to bind easily to the template at the
annealing temperature.
2. Primer melting temperature (Tm): Primers with melting
temperatures in the range of 52-58oC generally gives the best
results. The GC content of the sequence gives a fair indication of
the primer Tm. The two primers should be prepared in such a way
that their Tm difference should not be more than 2°C otherwise it
will result in poor annealing efficiency. Tm can be calculated by
the following formula:
10. Analysis of PCR results:
Once PCR cycle is complete, the amplified product is loaded in
the agarose gel and observed after ethidium bromide staining
under UV light source. A water blank reaction is included to
monitor the cross contaminating DNA source as template.
The percentage of agarose gel depends on the size of DNA to be
visualized. Generally 0.8-1% agarose gel is used for analyzing
0.5-5 kb amplified DNA while a DNA of larger size or genomic
DNA is visualized in gel as low as 0.5%.
11.
12. APPLICATIONS OF PCR
PCR in human medicine:
PCR technology has become an essential research and diagnostic
tool for improving human health and quality of life. It allows the
detection of infectious organisms just from one cell by amplifying
specific region of the genetic material. Some important areas in
medical research where PCR technology is employed include the
following:
PCR in infectious disease: PCR technology has become the basis for a broad
spectrum of clinical diagnostic tests for various infectious agents, including
viruses and bacteria (Figure 16.2). Besides detecting the presence of
pathogens, PCR allows us to quantify the amount of pathogens present in
patient‟s blood and this helps to monitor the progression of infection or
response to drug treatment. PCR has enabled the development of diagnostic
tests for many diseases such as, HIV-1, Hepatitis B and C viruses, Human
Papillomavirus, Chlamydia trachomatis, Neisseria gonorrhoeae,
Cytomegalovirus, Mycobacterium tuberculosis.
13.
14. Applications of PCR
PCR and genetic testing: PCR technology has recently become
a powerful tool to detect disease associated gene to predict the
presence of heart disease and cancers. Knowledge of disease
associated gene in the person predisposed to these disorders have
a chance to control the problem much in advance.
PCR in plant science: There are various fields in plant science
which requires the use of PCR technology for its
accomplishment.
Plant species identification: PCR technique has also been
employed in identification of plant species using species and
group-specific primers targeting chloroplast DNA. These assays
allowed identification of plants based on size-specific amplicons.
plants belonging to the same family has close primer-binding sites
and hence same amplicons size while plants belonging to
different species and groups have different primer-binding sites
and hence will result in different amplicons size.
15. Applications of PCR
PCR in tissue culture: It is used in analysis of DNA and specific genes in plant
cells at different stages of re-generation during in vitro culture along with RAPD
(random amplification of polymorphic DNA) technology. The level of
polymorphism in regenerated plants could be revealed by these dual techniques.
PCR in veterinary parasitology: Owing to its rapidity and sensitivity as
compared to antibody-based diagnosis, PCR met its uses in almost all aspects of
biological work including veterinary clinical diagnosis. Some examples of PCR
applications in veterinary parasitology:
Aujeszky’s disease (pseudorabis) virus of pigs: This virus causes abortion and
mortality in piglets. This disease has a latent period where there is no symptom of
infection making it difficult to eradicate the disease completely. For this reason,
PCR is considered to be appropriate tool for detecting latent cases of Aujeszky‟s
disease.
Bovine leukemia virus (BLV): This virus causes enzootic bovine leukosis. PCR
assay for detection of BLV was developed in 1991.
16. PCR in Forensic applications:
The most common use of PCR in forensic applications includes:
(1) Criminal investigation: A sample of blood, hair root or
tissue left in the crime scene can be used to identify a person
using PCR by comparing the DNA of the crime scene with that of
suspect or with DNA database of earlier convicts.
Evidence from decades-old crimes can be tested, confirming or
defending the people originally convicted.