Types of PCR include allele-specific PCR, which uses primers targeting single nucleotide polymorphisms to detect specific alleles. Assembly PCR artificially synthesizes long DNA sequences from overlapping oligonucleotides. Asymmetric PCR preferentially amplifies one DNA strand. Colony PCR screens bacterial clones. Hot-start PCR reduces nonspecific amplification. Inverse PCR amplifies known internal sequences after circularizing and digesting DNA. Ligation-mediated PCR uses linkers to amplify target fragments. Methylation-specific PCR identifies DNA methylation patterns. Long PCR amplifies over 5kb. Miniprimer PCR uses very short primers. Multiplex PCR targets multiple genes. Nested PCR uses two primer sets for increased sensitivity. Quantitative PCR measures starting DNA amounts
In this ppt, the various types of PCR such as real time PCR, Reverse transcription PCR, multiplex PCR, ligation chain PCR, nested PCR which is applied in diagnosis of diseases, identification of genetic disorders, determination of polymorphism and also in DNA fingerprinting analysis are described.
In this ppt, the various types of PCR such as real time PCR, Reverse transcription PCR, multiplex PCR, ligation chain PCR, nested PCR which is applied in diagnosis of diseases, identification of genetic disorders, determination of polymorphism and also in DNA fingerprinting analysis are described.
A detailed description about the basic steps involved in the - PCR - Polymerase Chain Reaction, its applications,its limitations and steps to overcome it.
Introduction to real-Time Quantitative PCR (qPCR) - Download the slidesQIAGEN
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.
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
A detailed description about the basic steps involved in the - PCR - Polymerase Chain Reaction, its applications,its limitations and steps to overcome it.
Introduction to real-Time Quantitative PCR (qPCR) - Download the slidesQIAGEN
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.
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
Lecture ON Polymerase Chain Reaction.
The polymerase chain reaction (PCR) is a powerful core molecular biology technique - Sometimes called "molecular photocopying. • Developed by Kary Mullis in 1985.
• It is an efficient and rapid in vitro method for enzymatic amplification of specific DNA or RNA sequences from nucleic acids of various sources. •
It generates microgram (µg) quantities of DNA copies (up to billion copies) of the desired DNA (or RNA) segment.
A simple PCR reaction consists of
i. A DNA preparation containing the desired segment to be amplified.
ii. A set of synthetic oligonucleotide primers that flank the target DNA
sequence, of about 20 bases long, specific, i.e., complementary.
iii. A thermostable DNA polymerase e.g., Taq isolated from the
bacterium Thermus acquaticus, Pfu – Pyrococcus furiosus and Vent
from Thermococcus litoralis. Pfu and Vent are more efficient than
Taq polymerase.
iv. Four deoxynucleoside triphosphate (dNTPs): TTP – thymidine
triphosphate, dCTP – deoxycyctidine triphosphate, dATP –
deoxyadenosine triphosphate and dGTP – deoxyguanosine
triphosphate
Food hygiene is more than cleanliness ......
Protecting food from risk of contamination, including harmful bacteria, poison and other foreign bodies.
Preventing any bacteria present multiplying to an extent which would result in the illness of consumers or the early spoilage of the food.
Destroying any harmful bacteria in the food by thorough cooking
or processing.
Discarding unfit or contaminated food.
T-Cell Activation
• Concept of immune response
• T cell-mediated immune response
• B cell-mediated immune response
I. Concept of immune response
• A collective and coordinated response to the introduction of foreign substances in an individual mediated by the cells and molecules in the immune system.
II. T cell-mediated immune response
• Cell-mediated immunity is the arm of the adaptive immune response whose role is to combat infection of intracellular pathogens, such as intracellular bacteria (mycobacteria, listeria monocytogens), viruses, protozoa, etc.
Major Histocompatibility Complex
MHC:
• Major Histocompatibility Complex
– Cluster of genes found in all mammals
– Its products play role in discriminating self/non-self
– Participant in both humoral and cell-mediated immunity
• MHC Act As Antigen Presenting Structures
• In Human MHC Is Found On Chromosome 6
– Referred to as HLA complex
• In Mice MHC Is Found On Chromosome 17
– Referred to as H-2 complex
• Genes Of MHC Organized In 3 Classes
– Class I MHC genes
• Glycoproteins expressed on all nucleated cells
• Major function to present processed Ags to TC
– Class II MHC genes
• Glycoproteins expressed on macrophages, B-cells, DCs
• Major function to present processed Ags to TH
– Class III MHC genes
• Products that include secreted proteins that have immune functions. Ex. Complement system, inflammatory molecules
Antigen Processing and Presentation MID
Antigens and “foreignness”
• Antigens (or, more properly, immunogens) have a series of features which confer immunogenicity.
• One of these features is “foreignness.”
• So, we can infer that – most often – antigens – ultimately – originate externally.
• (There are exceptions, of course. Some cells become transformed by disease [e. g., cancer] or by aging. In such instances, the antigens have an internal origin.)
Extinction of a particular animal or plant species occurs when there are no more individuals of that species alive anywhere in the world - the species has died out. This is a natural part of evolution. But sometimes extinctions happen at a much faster rate than usual. Natural Causes of Extinction.
Difference between In-Situ and Ex-Situ conservation
Conservation of biodiversity and genetic resources helps protect, maintain and recover endangered animal and plant species. There are mainly two strategies for the conservation of wildlife: In-situ conservation and Ex-situ conservation. Although, both the strategies aim to maintain and recover endangered species, they are different from each other. Let us see how they differ from each other!
Evolution Of Bacteria
Bacteria have existed from very early in the history of life on Earth. Bacteria fossils discovered in rocks date from at least the Devonian Period (419.2 million to 358.9 million years ago), and there are convincing arguments that bacteria have been present since early Precambrian time, about 3.5 billion years ago. Bacteria were widespread on Earth at least since the latter part of the Paleoproterozoic, roughly 1.8 billion years ago, when oxygen appeared in the atmosphere as a result of the action of the cyanobacteria. Bacteria have thus had plenty of time to adapt to their environments and to have given rise to numerous descendant forms.
Impact of Environment on Loss of Genetic Diversity and Speciation
Genetic variation describes naturally occurring genetic differences among individuals of the same species. This variation permits flexibility and survival of a population in the face of changing environmental circumstances. Consequently, genetic variation is often considered an advantage, as it is a form of preparation for the unexpected. But how does genetic variation increase or decrease? And what effect do fluctuations in genetic variation have on populations over time?
GENE ENVIRONMENT INTERACTION
Subtle differences in one person’s genes can cause them to respond differently to the same environmental exposure as another person. As a result, some people may develop a disease after being exposed to something in the environment while others may not.
As scientists learn more about the connection between genes and the environment, they pursue new approaches for preventing and treating disease that consider individual genetic codes.
How to store food in hot
The Good News
To maximize benefit of preservation, keep your food as fresh as possible for as long as possible. You can do this, even in the heat, by creating a “cooler” made from two basic terra cotta pots, one larger than the other. Put the smaller pot in the larger one, fill the gap with sand, and saturate the sand with water. Then cover it with a cloth. To add additional insulation from the heat, bury the pot up to its rim. The evaporation of moisture from the wet sand will cool the air around the food and help keep it fresh.
What is IUPAC naming?
In order to give compounds a name, certain rules must be followed. When naming organic compounds, the IUPAC (International Union of Pure and Applied Chemistry) nomenclature (naming scheme) is used. This is to give consistency to the names. It also enables every compound to have a unique name, which is not possible with the common names used (for example in industry). We will first look at some of the steps that need to be followed when naming a compound, and then try to apply these rules to some specific examples.
IUPAC Nomenclature
IUPAC nomenclature uses the longest continuous chain of carbon atoms to determine the basic root name of the compound. The root name is then modified due to the presence of different functional groups which replace hydrogen or carbon atoms in the parent structure.
Hybridization describes the bonding atoms from an atom's point of view. For a tetrahedral coordinated carbon (e.g. methane CH4), the carbon should have 4 orbitals with the correct symmetry to bond to the 4 hydrogen atoms.
INTRODUCTION:
Hybrid Orbitals
Developed by Linus Pauling, the concept of hybrid orbitals was a theory created to explain the structures of molecules in space. The theory consists of combining atomic orbitals (ex: s,p,d,f) into new hybrid orbitals (ex: sp, sp2, sp3).
1. Why Firefly give light during night?
2. Why atomic mass and Atomic numbers are given to elements ?
3. Why elements have been characterized and classified into different groups?
4. What is the transition of elements and what they play their role in elements stability?
Connector Corner: Automate dynamic content and events by pushing a buttonDianaGray10
Here is something new! In our next Connector Corner webinar, we will demonstrate how you can use a single workflow to:
Create a campaign using Mailchimp with merge tags/fields
Send an interactive Slack channel message (using buttons)
Have the message received by managers and peers along with a test email for review
But there’s more:
In a second workflow supporting the same use case, you’ll see:
Your campaign sent to target colleagues for approval
If the “Approve” button is clicked, a Jira/Zendesk ticket is created for the marketing design team
But—if the “Reject” button is pushed, colleagues will be alerted via Slack message
Join us to learn more about this new, human-in-the-loop capability, brought to you by Integration Service connectors.
And...
Speakers:
Akshay Agnihotri, Product Manager
Charlie Greenberg, Host
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
Key Trends Shaping the Future of Infrastructure.pdfCheryl Hung
Keynote at DIGIT West Expo, Glasgow on 29 May 2024.
Cheryl Hung, ochery.com
Sr Director, Infrastructure Ecosystem, Arm.
The key trends across hardware, cloud and open-source; exploring how these areas are likely to mature and develop over the short and long-term, and then considering how organisations can position themselves to adapt and thrive.
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.
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.
Search and Society: Reimagining Information Access for Radical FuturesBhaskar Mitra
The field of Information retrieval (IR) is currently undergoing a transformative shift, at least partly due to the emerging applications of generative AI to information access. In this talk, we will deliberate on the sociotechnical implications of generative AI for information access. We will argue that there is both a critical necessity and an exciting opportunity for the IR community to re-center our research agendas on societal needs while dismantling the artificial separation between the work on fairness, accountability, transparency, and ethics in IR and the rest of IR research. Instead of adopting a reactionary strategy of trying to mitigate potential social harms from emerging technologies, the community should aim to proactively set the research agenda for the kinds of systems we should build inspired by diverse explicitly stated sociotechnical imaginaries. The sociotechnical imaginaries that underpin the design and development of information access technologies needs to be explicitly articulated, and we need to develop theories of change in context of these diverse perspectives. Our guiding future imaginaries must be informed by other academic fields, such as democratic theory and critical theory, and should be co-developed with social science scholars, legal scholars, civil rights and social justice activists, and artists, among others.
"Impact of front-end architecture on development cost", Viktor TurskyiFwdays
I have heard many times that architecture is not important for the front-end. Also, many times I have seen how developers implement features on the front-end just following the standard rules for a framework and think that this is enough to successfully launch the project, and then the project fails. How to prevent this and what approach to choose? I have launched dozens of complex projects and during the talk we will analyze which approaches have worked for me and which have not.
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.
Let's dive deeper into the world of ODC! Ricardo Alves (OutSystems) will join us to tell all about the new Data Fabric. After that, Sezen de Bruijn (OutSystems) will get into the details on how to best design a sturdy architecture within ODC.
GraphRAG is All You need? LLM & Knowledge GraphGuy Korland
Guy Korland, CEO and Co-founder of FalkorDB, will review two articles on the integration of language models with knowledge graphs.
1. Unifying Large Language Models and Knowledge Graphs: A Roadmap.
https://arxiv.org/abs/2306.08302
2. Microsoft Research's GraphRAG paper and a review paper on various uses of knowledge graphs:
https://www.microsoft.com/en-us/research/blog/graphrag-unlocking-llm-discovery-on-narrative-private-data/
FIDO Alliance Osaka Seminar: Passkeys at Amazon.pdf
Different types of PCR
1. 1
Types of PCR
1. Allele-specific PCR:
A diagnostic or cloning technique which is based on single-nucleotide polymorphisms (SNPs)
(single-base differences in DNA). It requires prior knowledge of a DNA sequence, including
differences between alleles, and uses primers whose 3' ends encompass the SNP. PCR
amplification under stringent conditions is much less efficient in the presence of a mismatch
between template and primer, so successful amplification with an SNP-specific primer signals
presence of the specific SNP in a sequence.
2. Assembly PCR or Polymerase Cycling Assembly (PCA):
Artificial synthesis of long DNA sequences by performing PCR on a pool of long
oligonucleotides with short overlapping segments. The oligonucleotides alternate between sense
and antisense directions, and the overlapping segments determine the order of the PCR
fragments, thereby selectively producing the final long DNA product.
2. 2
3. Asymmetric PCR:
Preferentially amplifies one DNA strand in a double-stranded DNA template. It is used in
sequencing and hybridization probing where amplification of only one of the two complementary
strands is required. PCR is carried out as usual, but with a great excess of the primer for the
strand targeted for amplification. Because of the slow (arithmetic) amplification later in the
reaction after the limiting primer has been used up, extra cycles of PCR are required. A recent
modification on this process, known as Linear-After-The-Exponential-PCR (LATE-PCR), uses a
limiting primer with a higher melting temperature (Tm) than the excess primer to maintain
reaction efficiency as the limiting primer concentration decreases mid-reaction.
4. Colony PCR :
The screening of bacterial (E.Coli) or yeast clones for correct ligation or plasmid products.
Selected colonies of bacteria or yeast are picked with a sterile toothpick or pipette tip from a
growth (agarose) plate. This is then inserted into the PCR master mix or pre-inserted into
autoclaved water. PCR is then conducted to determine if the colony contains the DNA fragment
or plasmid of interest.
5. Hot-start PCR:
A technique that reduces non-specific amplification during the initial set up stages of the PCR. It
may be performed manually by heating the reaction components to the melting temperature (e.g.,
95°C) before adding the polymerase. Specialized enzyme systems have been developed that
inhibit the polymerase's activity at ambient temperature, either by the binding of an antibody or
by the presence of covalently bound inhibitors that only dissociate after a high-temperature
activation step. Hot-start/cold-finish PCR is achieved with new hybrid polymerases that are
inactive at ambient temperature and are instantly activated at elongation temperature.
6. Inverse PCR Steps
Target DNA is lightly cut into smaller fragments of several kilobases by restriction
endonuclease digestion.
3. 3
Self-ligation is induced under low concentrations causing the phosphate backbone to
reform. This gives a circular DNA ligation product.
Target DNA is then restriction digested with a known endonuclease. This generates a cut
within the known internal sequence generating a linear product with known terminal
sequences. This can now be used for PCR (polymerase chain reaction).
Standard PCR is conducted with primers complementary to the now known internal
sequences.
7. Ligation-mediated PCR :
Uses small DNA oligonucleotide 'linkers' (or adaptors) that are first ligated to fragments of the
target DNA. PCR primers that anneal to the linker sequences are then used to amplify the target
fragments. This method is deployed for DNA sequencing, genome walking, and DNA
footprinting A related technique is Amplified fragment length polymorphism, which generates
diagnostic fragments of a genome.
8. Methylation-specific PCR (MSP):
Is used to identify patterns of DNA methylation at cytosine-guanine (CpG) islands in genomic
DNA.Target DNA is first treated with sodium bisulfite, which converts unmethylated cytosine
bases to uracil, which is complementary to adenosine in PCR primers. Two amplifications are
then carried out on the bisulfite-treated DNA: One primer set anneals to DNA with cytosines
4. 4
(corresponding to methylated cytosine), and the other set anneals to DNA with uracil
(corresponding to unmethylated cytosine). MSP used in Q-PCR provides quantitative
information about the methylation state of a given CpG island.
9. Long PCR:
Is a PCR is which extended or longer than standard PCR, meaning over 5 kilobases (frequently over 10
kb). Long PCR is usually only useful if it is accurate. Thus, special mixtures of proficient polymerases
along with accurate polymerases such as Pfu are often mixed together. Applications of Long PCR Long
PCR is often used to clone larger genes or large segments of DNA which standard PCR cannot.
10. Miniprimer PCR:
Uses a thermostable polymerase (S-Tbr) that can extend from short primers ("smalligos") as short as 9 or
10 nucleotides. This method permits PCR targeting to smaller primer binding regions, and is used to
amplify conserved DNA sequences,such as the 16S (or eukaryotic 18S) rRNA gene.
11. Multiplex-PCR:
Consists of multiple primer sets within a single PCR mixture to produce amplicons of varying
sizes that are specific to different DNA sequences. By targeting multiple genes at once,
additional information may be gained from a single test run that otherwise would require several
times the reagents and more time to perform. Annealing temperatures for each of the primer sets
must be optimized to work correctly within a single reaction, and amplicon sizes, i.e., their base
pair length, should be different enough to form distinct bands when visualized by gel
electrophoresis.
5. 5
12. Nested PCR:
This PCR increases the sensitivity
Two sets of primers,
A double process of amplification .
The first set of primers allow a first amplification. The product of this PCR is subjected
to a second PCR using the second set of primers.
Primers used in the second PCR are specific to an internal amplified sequence in the first
PCR. specificity of the first PCR product is verified with the second one.
13. Quantitative PCR (Q-PCR):
Used to measure the quantity of a PCR product (commonly in real-time). It quantitatively
measures starting amounts of DNA, cDNA or RNA. Q-PCR is commonly used to determine
whether a DNA sequence is present in a sample and the number of its copies in the sample.
Quantitative real-time PCR has a very high degree of precision. QRT-PCR methods use
fluorescent dyes, such as Sybr Green, EvaGreen or fluorophore-containing DNA probes, such as
TaqMan, to measure the amount of amplified product in real time. It is also sometimes
abbreviated to RT-PCR (Real Time PCR) or RQ-PCR. QRT-PCR or RTQ-PCR are more
appropriate contractions, since RT-PCR commonly refers to reverse transcription PCR (see
below), often used in conjunction with Q-PCR.
6. 6
14. Reverse Transcription PCR (RT-PCR):
For amplifying DNA from RNA. Reverse transcriptase reverse transcribes RNA into cDNA,
which is then amplified by PCR. RT-PCR is widely used in expression profiling, to determine
the expression of a gene or to identify the sequence of an RNA transcript, including transcription
start and termination sites. If the genomic DNA sequence of a gene is known, RT-PCR can be
used to map the location of exons and introns in the gene. The 5' end of a gene (corresponding to
the transcription start site) is typically identified by RACE-PCR (Rapid Amplification of cDNA
Ends).
15. Touchdown PCR (Step-down PCR):
A variant of PCR that aims to reduce nonspecific background by gradually lowering the
annealing temperature as PCR cycling progresses. The annealing temperature at the initial cycles
is usually a few degrees (3-5°C) above the Tm of the primers used, while at the later cycles, it is
a few degrees (3-5°C) below the primer Tm. The higher temperatures give greater specificity for
primer binding, and the lower temperatures permit more efficient amplification from the specific
products formed during the initial cycles.
7. 7
16. Universal Fast Walking:
For genome walking and genetic fingerprinting using a more specific 'two-sided' PCR than
conventional 'one-sided' approaches (using only one gene-specific primer and one general primer
- which can lead to artefactual 'noise') by virtue of a mechanism involving lariat structure
formation. Streamlined derivatives of UFW are LaNe RAGE (lariat-dependent nested PCR for
rapid amplification of genomic DNA ends), 5'RACE LaNe and 3'RACE LaNe.
17. Variable Number of Tandem Repeats (VNTR) PCR :
Targets areas of the genome that exhibit length variation. The analysis of the genotypes of the
sample usually involves sizing of the amplification products by gel electrophoresis. Analysis of
smaller VNTR segments known as Short Tandem Repeats (or STRs) is the basis for DNA
Fingerprinting databases such as CODIS.