Sanger sequencing: the most popular method first devised by Fred Sanger and colleagues in the mid-1970
DNA to be sequenced serves as a template for DNA synthesis.
A DNA primer is designed to be a starting point for DNA synthesis by DNA polymerase on the strand of DNA to be sequenced.
Four individual DNA synthesis reactions are performed.
The four reactions include normal A, G, C, and T deoxynucleotide triphosphates (dNTPs), and each contains a low level of one of four dideoxynucleotide triphosphates (ddNTPs): ddATP, ddGTP, ddCTP, or ddTTP.
Sanger sequencing: the most popular method first devised by Fred Sanger and colleagues in the mid-1970
DNA to be sequenced serves as a template for DNA synthesis.
A DNA primer is designed to be a starting point for DNA synthesis by DNA polymerase on the strand of DNA to be sequenced.
Four individual DNA synthesis reactions are performed.
The four reactions include normal A, G, C, and T deoxynucleotide triphosphates (dNTPs), and each contains a low level of one of four dideoxynucleotide triphosphates (ddNTPs): ddATP, ddGTP, ddCTP, or ddTTP.Sanger sequencing: the most popular method first devised by Fred Sanger and colleagues in the mid-1970
DNA to be sequenced serves as a template for DNA synthesis.
A DNA primer is designed to be a starting point for DNA synthesis by DNA polymerase on the strand of DNA to be sequenced.
Four individual DNA synthesis reactions are performed.
The four reactions include normal A, G, C, and T deoxynucleotide triphosphates (dNTPs), and each contains a low level of one of four dideoxynucleotide triphosphates (ddNTPs): ddATP, ddGTP, ddCTP, or ddTTP.
Sanger sequencing: the most popular method first devised by Fred Sanger and colleagues in the mid-1970
DNA to be sequenced serves as a template for DNA synthesis.
A DNA primer is designed to be a starting point for DNA synthesis by DNA polymerase on the strand of DNA to be sequenced.
Four individual DNA synthesis reactions are performed.
The four reactions include normal A, G, C, and T deoxynucleotide triphosphates (dNTPs), and each contains a low level of one of four dideoxynucleotide triphosphates (ddNTPs): ddATP, ddGTP, ddCTP, or ddTTP.
The starting template material is RNA not DNA ( as in PCR assays for the diagnosis of viral infections)
RNA cannot serve as a template for PCR, (RNA is not a substrate for the Taq DNA polymerases commonly utilised in PCR.) Therefore reverse transcription is combined with PCR to convert RNA into a complementary DNA (cDNA)) suitable for PCR
The first step in this procedure is to convert the RNA molecules into single-stranded complementary DNA (cDNA) (Figure 9.20). Once this preliminary step has been carried out, the PCR primers and Taq polymerase are added and the experiment proceeds exactly as in the standard technique
The starting template material is RNA not DNA ( as in PCR assays for the diagnosis of viral infections)
RNA cannot serve as a template for PCR, (RNA is not a substrate f
Polymerase chain reaction (PCR) is a technique in molecular biology used to
amplify (multiply) a single copy or a few copies of a piece of DNA, generating
thousands to millions of copies of that particular DNA sequence.
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 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.
Polymerase chain reaction is a technique used in molecular biology to amplify a single copy or a few copies of a segment of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence
Polymerase chain reaction (PCR)
Polymerase chain reaction (PCR) is a common laboratory technique used to make many copies (millions or billions) of a particular region of DNA.
Polymerase chain reaction (PCR) is a technique in molecular biology used to
amplify (multiply) a single copy or a few copies of a piece of DNA, generating
thousands to millions of copies of that particular DNA sequence.
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 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.
Polymerase chain reaction is a technique used in molecular biology to amplify a single copy or a few copies of a segment of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence
Polymerase chain reaction (PCR)
Polymerase chain reaction (PCR) is a common laboratory technique used to make many copies (millions or billions) of a particular region of DNA.
PCR (polymerase chain reaction) is a method to analyze a short sequence of DNA (or RNA) even in samples containing only minute quantities of DNA or RNA. PCR is used to reproduce (amplify) selected sections of DNA or RNA.
Polymerase chain reaction (PCR) is a technique used in molecular biology to amplify a single copy or a few copies of a segment of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. It is an easy, cheap, and reliable way to repeatedly replicate a focused segment of DNA, a concept which is applicable to numerous fields in modern biology and related sciences.
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
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
b pharmacy
pharmaceutical biotechnology
Polymerase chain reaction
History
Purpose
Components of PCR
Steps of PCR
Denaturation of DNA template
Annealing of primers
Extension of ds DNA molecules
Reaction Condition & Experimental Protocol
General PCR Protocol
Application
1- Biochemical and molecular basis of lung diseases .pptMohamed Afifi
Recognize the biochemical structure and function of pulmonary surfactant
Discuss biochemical basis of respiratory distress syndrome
List the differences between collagen and elastin.
Identify the biochemical basis of lung emphysema due to alpha one antitrypsin deficiency.
Outline the biochemical and molecular basis of cystic fibrosis
Mention the diagnosis and treatment of cystic fibrosis
Definition f microorganism
A microorganism, or microbe, is an organism of microscopic size, which may exist in its single-celled form or as a colony of cells
PCR (polymerase chain reaction) is a method to analyze a short sequence of DNA (or RNA) even in samples containing only minute quantities of DNA or RNA. PCR is used to reproduce (amplify) selected sections of DNA or RNA.
Polymerase chain reaction (PCR) is a technique used in molecular biology to amplify a single copy or a few copies of a segment of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. It is an easy, cheap, and reliable way to repeatedly replicate a focused segment of DNA, a concept which is applicable to numerous fields in modern biology and related sciences.
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
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
b pharmacy
pharmaceutical biotechnology
Polymerase chain reaction
History
Purpose
Components of PCR
Steps of PCR
Denaturation of DNA template
Annealing of primers
Extension of ds DNA molecules
Reaction Condition & Experimental Protocol
General PCR Protocol
Application
1- Biochemical and molecular basis of lung diseases .pptMohamed Afifi
Recognize the biochemical structure and function of pulmonary surfactant
Discuss biochemical basis of respiratory distress syndrome
List the differences between collagen and elastin.
Identify the biochemical basis of lung emphysema due to alpha one antitrypsin deficiency.
Outline the biochemical and molecular basis of cystic fibrosis
Mention the diagnosis and treatment of cystic fibrosis
Definition f microorganism
A microorganism, or microbe, is an organism of microscopic size, which may exist in its single-celled form or as a colony of cells
Fate of Absorbed Amino Acids
Absorbed amino acids from diet are mixed with the amino acids produced from
hydrolysis of body protein and those synthesized in the body to form a common
amino acid pool (about 100g). This pool is drawn upon for anabolism and for
catabolism of amino acids.
Anabolic Fate
-These include the synthesis of proteins e.g. tissue, milk, and plasma proteins,
enzymes, and some hormones.
-They also include the synthesis of other nitrogenous substances e.g. glutathione,
adrenaline, thyroxine, melanin, niacin, purines, pyrimidines, aminosugars, and the
nitrogenous bases of phospholipids.
Catabolic Fate
-Most catabolic reactions are preceded by cleavage of the amino acids into ammonia
and the carbon skeleton (usually in the form of an -keto acid).
-The ammonia is mostly converted to urea, which is excreted in urine.
-Little ammonia is also excreted in urine. The carbon skeleton m
Cells of the nervous system: Neurons & Glial cells
▫ Glial cells:
most abundant cells, are of 3 main types
A. Microglia: Phagocytic (immune) cells,
B. Astrocytes: Support & nutrition of the neurons
(provide neurons with lactate from glucose and
regulate the content of ECF by removing K+ &
excess neurotransmitters)
C. Oligodendrocytes: form myelin sheathe in CNS
Schwan cells form myelin sheaths in PNS
Ependymal cells: line brain & spinal cord cavities
using their cilia to allow for the circulation of the
1- metabolism of the brain (I) 2012-13.pdfMohamed Afifi
Cells of the nervous system: Neurons & Glial cells
▫ Neurons:
A neuron is Formed of:
Cell body:
▫ contains most of the cytoplasm & organelles
Cytoplasmic extensions:
▫ include an axon & many dendrites
Overvie
The starting template material is RNA not DNA ( as in PCR assays for the diagnosis of viral infections)
RNA cannot serve as a template for PCR, (RNA is not a substrate for the Taq DNA polymerases commonly utilised in PCR.) Therefore reverse transcription is combined with PCR to convert RNA into a complementary DNA (cDNA)) suitable for PCR
The first step in this procedure is to convert the RNA molecules into single-stranded complementary DNA (cDNA) (Figure 9.20). Once this preliminary step has been carried out, the PCR primers and Taq polymerase are added and the experiment proceeds exactly as in the standard technique
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Care must be taken to preserve protein structure and function after it is removed
from its natural environment where it was stable.
• pH – To prevent denaturation or loss of function, proteins are placed in buffered
solutions at or near their native pH.
• Temperature – Protein purification is normally carried at low temperature ~ 0°C.
while some proteins are thermally stable at high temperatures.
• Inhibition of proteases
• Retardation of microbes that can destroy proteins
Sodium azide is often used
Salting-In: Most globular proteins tend to become
increasingly soluble as the ionic strength is raised due to
the addition of salt. This phenomenon is known as saltingin of proteins.
• Salting-out: As the salt concentration increases, this
lead to diminishment of electrostatic attraction between
protein molecules by the presence of abundant salt ions.
This phenomenon is known as salting-out of proteins.
• The salt concentration at which protein precipitates differs
from one protein to another.
• Salting out is one of the most commonly used protein
purification procedures.
• Ammonium sulfate is the most commonly used reagent
- High solubility (3.9 M in water at 0 ºC)
- High ionic strength solution can be made (up to 23.5 in
water at 0 ºC)
There are four steps of ion-exchange chromatography:
1. Equilibration – stabilization of the ion-exchangers with oppositely
charged ions in the buffer. E.g: Na+Cl2. Sample application and wash (Protein bound to the ion-exchangers
remain attached while other gets removed during wash)
3. Elution – Removal of bound protein from the ion exchangers with the
help of increased concentration of elution buffer.
4. Regeneration – Preparing the ion exchangers for the next round of
protein purification
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Size-Exclusion (or molecular
exclusion) Chromatography
•Molecules are separated
according to differences in their
size as they pass through a
hydrophilic polymer
•Polymer beads composed of
cross-linked dextran (dextrose)
which is highly and uniformly
porous (like Swiss cheese)
•Large proteins come out first
(can’t fit in pores), small proteins
come out last (get stuck in the
por
There are four steps of ion-exchange chromatography:
1. Equilibration – stabilization of the ion-exchangers with oppositely
charged ions in the buffer. E.g: Na+Cl2. Sample application and wash (Protein bound to the ion-exchangers
remain attached while othe
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Empowering the Data Analytics Ecosystem: A Laser Focus on Value
The data analytics ecosystem thrives when every component functions at its peak, unlocking the true potential of data. Here's a laser focus on key areas for an empowered ecosystem:
1. Democratize Access, Not Data:
Granular Access Controls: Provide users with self-service tools tailored to their specific needs, preventing data overload and misuse.
Data Catalogs: Implement robust data catalogs for easy discovery and understanding of available data sources.
2. Foster Collaboration with Clear Roles:
Data Mesh Architecture: Break down data silos by creating a distributed data ownership model with clear ownership and responsibilities.
Collaborative Workspaces: Utilize interactive platforms where data scientists, analysts, and domain experts can work seamlessly together.
3. Leverage Advanced Analytics Strategically:
AI-powered Automation: Automate repetitive tasks like data cleaning and feature engineering, freeing up data talent for higher-level analysis.
Right-Tool Selection: Strategically choose the most effective advanced analytics techniques (e.g., AI, ML) based on specific business problems.
4. Prioritize Data Quality with Automation:
Automated Data Validation: Implement automated data quality checks to identify and rectify errors at the source, minimizing downstream issues.
Data Lineage Tracking: Track the flow of data throughout the ecosystem, ensuring transparency and facilitating root cause analysis for errors.
5. Cultivate a Data-Driven Mindset:
Metrics-Driven Performance Management: Align KPIs and performance metrics with data-driven insights to ensure actionable decision making.
Data Storytelling Workshops: Equip stakeholders with the skills to translate complex data findings into compelling narratives that drive action.
Benefits of a Precise Ecosystem:
Sharpened Focus: Precise access and clear roles ensure everyone works with the most relevant data, maximizing efficiency.
Actionable Insights: Strategic analytics and automated quality checks lead to more reliable and actionable data insights.
Continuous Improvement: Data-driven performance management fosters a culture of learning and continuous improvement.
Sustainable Growth: Empowered by data, organizations can make informed decisions to drive sustainable growth and innovation.
By focusing on these precise actions, organizations can create an empowered data analytics ecosystem that delivers real value by driving data-driven decisions and maximizing the return on their data investment.
Adjusting primitives for graph : SHORT REPORT / NOTESSubhajit Sahu
Graph algorithms, like PageRank Compressed Sparse Row (CSR) is an adjacency-list based graph representation that is
Multiply with different modes (map)
1. Performance of sequential execution based vs OpenMP based vector multiply.
2. Comparing various launch configs for CUDA based vector multiply.
Sum with different storage types (reduce)
1. Performance of vector element sum using float vs bfloat16 as the storage type.
Sum with different modes (reduce)
1. Performance of sequential execution based vs OpenMP based vector element sum.
2. Performance of memcpy vs in-place based CUDA based vector element sum.
3. Comparing various launch configs for CUDA based vector element sum (memcpy).
4. Comparing various launch configs for CUDA based vector element sum (in-place).
Sum with in-place strategies of CUDA mode (reduce)
1. Comparing various launch configs for CUDA based vector element sum (in-place).
As Europe's leading economic powerhouse and the fourth-largest hashtag#economy globally, Germany stands at the forefront of innovation and industrial might. Renowned for its precision engineering and high-tech sectors, Germany's economic structure is heavily supported by a robust service industry, accounting for approximately 68% of its GDP. This economic clout and strategic geopolitical stance position Germany as a focal point in the global cyber threat landscape.
In the face of escalating global tensions, particularly those emanating from geopolitical disputes with nations like hashtag#Russia and hashtag#China, hashtag#Germany has witnessed a significant uptick in targeted cyber operations. Our analysis indicates a marked increase in hashtag#cyberattack sophistication aimed at critical infrastructure and key industrial sectors. These attacks range from ransomware campaigns to hashtag#AdvancedPersistentThreats (hashtag#APTs), threatening national security and business integrity.
🔑 Key findings include:
🔍 Increased frequency and complexity of cyber threats.
🔍 Escalation of state-sponsored and criminally motivated cyber operations.
🔍 Active dark web exchanges of malicious tools and tactics.
Our comprehensive report delves into these challenges, using a blend of open-source and proprietary data collection techniques. By monitoring activity on critical networks and analyzing attack patterns, our team provides a detailed overview of the threats facing German entities.
This report aims to equip stakeholders across public and private sectors with the knowledge to enhance their defensive strategies, reduce exposure to cyber risks, and reinforce Germany's resilience against cyber threats.
Data Centers - Striving Within A Narrow Range - Research Report - MCG - May 2...pchutichetpong
M Capital Group (“MCG”) expects to see demand and the changing evolution of supply, facilitated through institutional investment rotation out of offices and into work from home (“WFH”), while the ever-expanding need for data storage as global internet usage expands, with experts predicting 5.3 billion users by 2023. These market factors will be underpinned by technological changes, such as progressing cloud services and edge sites, allowing the industry to see strong expected annual growth of 13% over the next 4 years.
Whilst competitive headwinds remain, represented through the recent second bankruptcy filing of Sungard, which blames “COVID-19 and other macroeconomic trends including delayed customer spending decisions, insourcing and reductions in IT spending, energy inflation and reduction in demand for certain services”, the industry has seen key adjustments, where MCG believes that engineering cost management and technological innovation will be paramount to success.
MCG reports that the more favorable market conditions expected over the next few years, helped by the winding down of pandemic restrictions and a hybrid working environment will be driving market momentum forward. The continuous injection of capital by alternative investment firms, as well as the growing infrastructural investment from cloud service providers and social media companies, whose revenues are expected to grow over 3.6x larger by value in 2026, will likely help propel center provision and innovation. These factors paint a promising picture for the industry players that offset rising input costs and adapt to new technologies.
According to M Capital Group: “Specifically, the long-term cost-saving opportunities available from the rise of remote managing will likely aid value growth for the industry. Through margin optimization and further availability of capital for reinvestment, strong players will maintain their competitive foothold, while weaker players exit the market to balance supply and demand.”
Opendatabay - Open Data Marketplace.pptxOpendatabay
Opendatabay.com unlocks the power of data for everyone. Open Data Marketplace fosters a collaborative hub for data enthusiasts to explore, share, and contribute to a vast collection of datasets.
First ever open hub for data enthusiasts to collaborate and innovate. A platform to explore, share, and contribute to a vast collection of datasets. Through robust quality control and innovative technologies like blockchain verification, opendatabay ensures the authenticity and reliability of datasets, empowering users to make data-driven decisions with confidence. Leverage cutting-edge AI technologies to enhance the data exploration, analysis, and discovery experience.
From intelligent search and recommendations to automated data productisation and quotation, Opendatabay AI-driven features streamline the data workflow. Finding the data you need shouldn't be a complex. Opendatabay simplifies the data acquisition process with an intuitive interface and robust search tools. Effortlessly explore, discover, and access the data you need, allowing you to focus on extracting valuable insights. Opendatabay breaks new ground with a dedicated, AI-generated, synthetic datasets.
Leverage these privacy-preserving datasets for training and testing AI models without compromising sensitive information. Opendatabay prioritizes transparency by providing detailed metadata, provenance information, and usage guidelines for each dataset, ensuring users have a comprehensive understanding of the data they're working with. By leveraging a powerful combination of distributed ledger technology and rigorous third-party audits Opendatabay ensures the authenticity and reliability of every dataset. Security is at the core of Opendatabay. Marketplace implements stringent security measures, including encryption, access controls, and regular vulnerability assessments, to safeguard your data and protect your privacy.
2. RESPIRATION
(1) Pulmonary ventilation
(2) External respiration
Respiratory gas transport
(3)
Internal respiration
(4)
Breathing
: means
movement
of air In and
Outof thebody
Oxygen loading and
carbondioxide
loading
Transportation of gasesby blood stream
Exchange between capillary and body ce
3. (1) BREATHING (PULMONARY VENTILATION)
Inspiration: Diaphragm flattens creates a vacuum pulling
air into the lungs
Expiration: Muscles relax and push air out of the lungs
7. Pulmonary Function Tests
Definition:
Pulmonary function tests (PFTs) are noninvasive tests that show how well the
lungs are working. The tests measure lung volume, capacity, rates of flow, and
gas exchange. This information can help your healthcare provider diagnose
and decide the treatment of certain lung disorders
Types of PFTs
1-Spirometry measures the rate of air flow and estimates lung size. For
this test, you will breathe multiple times, with regular and maximal effort,
through a tube that is connected to a computer. Some people feel lightheaded
or tired from the required breathing effort.
.
8. 2-Lung volume tests are the most accurate
way to measure how much air your lungs can
hold.
3-Lung diffusion capacity assesses how well
oxygen gets into the blood from the air you
breathe.
3-Pulse oximetry estimates oxygen levels in
your blood
4-Arterial blood gas tests
5-Fractional exhaled nitric oxide tests
11. •Arterial blood gas tests
•Arterial blood gas tests directly measure the levels of gases, such as oxygen and
carbon dioxide, in blood.
Collection and handling of arterial blood gases:
1)The specimen for blood gases and pH should be arterial or arterialized
capillary blood
2)All air bubbles should be removed.
3)Air contamination will reduce the CO2 and increases the O2 in the sample
due to the difference in the PO2 and PCO2 tension of these gases in the
atmosphere.
4)Use the correct amount of heparin (0.05 mg heparin/ml blood).
5)The specimen must be placed in ice water until analysis or examined
immediately.
12. Importance of pulmonary function tests?
•They are important for detection of :
•Allergies
•Respiratory infections
•Trouble breathing from injury to the chest or a recent surgery
•Chronic lung conditions, such as asthma, bronchiectasis,
emphysema, or chronic bronchitis
•Asbestosis, a lung disease caused by inhaling asbestos fibers
•Restrictive airway problems from scoliosis, tumors, or
inflammation or scarring of the lungs
•Sarcoidosis, a disease that causes lumps of inflammatory cells
around organs, such as the liver, lungs, and spleen
•Scleroderma, a disease that causes thickening and hardening of
connective tissue
13. •the 5' to 3' exonuclease activity of
the Taq polymerase degrades the probe that has
annealed to the template.
• Degradation of the probe releases the
fluorophore from it and breaks the proximity to
the quencher, thus relieving the quenching effect
and allowing fluorescence of the fluorophore.
•Hence, fluorescence detected in the quantitative
PCR thermal cycler is directly proportional to the
fluorophore released and the amount of DNA
template present in the PCR.
14. 1). In this plot, the number of
PCR cycles is shown on the
x-axis, and the fluorescence
from the amplification
reaction, which is
proportional to the amount of
amplified product in the tube,
is shown on the y-axis.
15. Initially, fluorescence remains at background levels, and increases in
fluorescence are not detectable (cycles 1–18, Figure 1) even though product
accumulates exponentially.
Eventually, enough amplified product accumulates to yield a detectable
fluorescence signal.
The cycle number at which this occurs is called the quantification cycleCq, or.
Threshold cycle Ct
, The Threshold Cycle (Ct) or the quantification cycle (Cq): is the number
of cycles required for the fluorescent signal to cross the threshold (exceeds
background level), or it is the PCR cycle at which the amplification curve
intercepts with the threshold line
16. The Cq of a reaction is determined mainly by the amount of template present at
the start of the amplification reaction.
If a large amount of template is present at the start of the reaction, relatively
few amplification cycles will be required to accumulate enough product to give a
fluorescence signal above background. Thus, the reaction will have a low, or
early, Cq.
In contrast, if a small amount of template is present at the start of the reaction,
more amplification cycles will be required for the fluorescence signal to rise
above background. Thus, the reaction will have a high, or late, Cq.
This relationship forms the basis for the quantitative aspect of real-time PCR.
17. Blue sample Ct value=23. Green sample Ct value=28. Therefore the blue
sample contained 32 (25 ) times more of the gene of interest than the green
sample.
18. comparison between real time PCR and traditional PCR
•Traditional PCR: uses agarose gel
for detection of PCR amplification at
the plateau phase or end-point of the
PCR reaction.
•If measurements were taken at the
plateau phase, the data would not truly
represent the initial amounts of starting
target material.
•Real-Time PCR provides fast, precise
and accurate results. It is designed to
collect data as the reaction is
proceeding (exponential phase ),
which is more accurate for DNA and
RNA quantitation and does not require
post PCR methods
19. 2-Reverse Transcription Quantitative PCR (RT-qPCR)
• The starting template material is RNA not DNA ( as in PCR assays for the
diagnosis of viral infections)
• RNA cannot serve as a template for PCR, (RNA is not a substrate for the
Taq DNA polymerases commonly utilised in PCR.) Therefore reverse
transcription is combined with PCR to convert RNA into a complementary
DNA (cDNA)) suitable for PCR
• The first step in this procedure is to convert the RNA molecules into single-
stranded complementary DNA (cDNA) (Figure 9.20). Once this preliminary
step has been carried out, the PCR primers and Taq polymerase are
added and the experiment proceeds exactly as in the standard technique
20.
21. The RT reaction
• one-step procedure: The reverse transcription process and the PCR
can be performed at the same time where all the components for RT and
PCR are present from the start, i.e., RNA, primers, dNTP’s, PCR buffer,
RT and Taq enzymes (or an enzyme that can perform both functions).
Two steps procedure: involves creating cDNA first by means of a
separate reverse transcription reaction and then adding the cDNA to the
PCR reaction (more preferred better control the assay and to perform a
reaction titration)
22. DNA sequencing:
• Def: DNAsequencingis the processof determining the sequence of nucleotide
bases (As,T
s,Cs,and Gs)in apiece of DNA
•There are several procedures for DNA sequencing:
1. Maxam and Gilbert Method: method based on chemical
modification of DNA and subsequent cleavage at specific bases
2. ChainTermination(Sanger)Sequencing:A modified DNA replication
reaction. Growing chains are terminated by dideoxynucleotides
3. Pyro sequencing : new generation method that detects light
emitted during the sequential addition of nucleotides during the
synthesis of a complementary strand of DNA.
23. Sanger sequencing: the most popular method first devised by Fred
Sanger and colleagues in the mid-1970
•DNA to be sequenced serves as a template for DNA synthesis.
•A DNA primer is designed to be a starting point for DNA synthesis by
DNA polymerase on the strand of DNA to be sequenced.
•Four individual DNA synthesis reactions are performed.
•The four reactions include normal A, G, C, and T deoxynucleotide
triphosphates (dNTPs), and each contains a low level of one of four
dideoxynucleotide triphosphates (ddNTPs): ddATP, ddGTP, ddCTP, or
ddTTP.
24. •The four reactions can be named A, G, C
and T, according to which of the four
ddNTPs was included.
•When a ddNTP is incorporated into a
chain of nucleotides, synthesis terminates.
This is because the ddNTP molecule lacks
a 3' hydroxyl group, which is required to
form a link with the next nucleotide in the
chain.
•Since the ddNTPs are randomly
incorporated, synthesis terminates at many
different positions for each reaction
25. •Following synthesis, the products of the A, G, C, and T reactions are
individually loaded into four lanes of a single gel and separated using gel
electrophoresis
•The bands of the gel are detected, and then the sequence is read from
the bottom of the gel to the top, including bands in all four lanes.
For instance, if the lowest band
across all four lanes appears in the A
reaction lane, then the first
nucleotide in the sequence is A.
Then if the next band from bottom to
top appears in the T lane, the
second nucleotide in the sequence
is T, and so on.