In this slidedeck, we reveal how to get the most from your FFPE samples. We discuss variability in quantity and purity of DNA purified from FFPE samples manually or with automated procedures, assessed by different quantification and quality control methods. You can also learn more about the QIAxpert system and how it can help you gain reliable quantification of FFPE samples.
Cancer Research & the Challenges of FFPE Samples – An IntroductionQIAGEN
A cascade of complex genetic and epigenetic changes regulate tumor formation and progression. Gene expression analyses can shed light on these changes at a molecular level and identify the key genes and associated pathways involved in cancer. Often the samples used in cancer research are FFPE samples, which pose a significant challenge in terms of nucleic acid quality. The quality of nucleic acids extracted from FFPE samples depends on a number of factors, including how the samples were handled before, during and after fixation and embedding.
Dr. Vishwadeepak Tripathi describes the variability of sample purification from FFPE samples – in particular, samples to be used in cancer research. What are the challenges and solutions, and what quality control approach can ensure credible results? This webinar will focus on sample purification and the quality control of FFPE samples and compare different automated purification procedures.
Critical Factors for Successful Real-Time PCR: Multiplex PCRQIAGEN
Multiplex end-point PCR is a powerful tool for genotyping and many other applications. QIAGEN’s multiplex PCR chemistry is optimized for reliable amplification of many different templates with high variability in copy numbers. Thus it enables very quick establishment of a new lab routine and instant success for your multiplex PCR strategy.
There is a set of critical factors which we recommend to be regarded for planning and performing this kind of PCR. These will be discussed in detail in the webinar. Additionally, our multiplex PCR chemistry has recently been gaining increasing popularity among scientists who are utilizing it for their next-generation sequencing workflows.
Quality control lecture CPath master 2014 Ain ShamsAdel Elazab Elged
Basics of quality management or assurance program detailing values of internal quality control material analysis and interpretation and external quality control or proficiency testing programs in medical laboratories
Cancer Research & the Challenges of FFPE Samples – An IntroductionQIAGEN
A cascade of complex genetic and epigenetic changes regulate tumor formation and progression. Gene expression analyses can shed light on these changes at a molecular level and identify the key genes and associated pathways involved in cancer. Often the samples used in cancer research are FFPE samples, which pose a significant challenge in terms of nucleic acid quality. The quality of nucleic acids extracted from FFPE samples depends on a number of factors, including how the samples were handled before, during and after fixation and embedding.
Dr. Vishwadeepak Tripathi describes the variability of sample purification from FFPE samples – in particular, samples to be used in cancer research. What are the challenges and solutions, and what quality control approach can ensure credible results? This webinar will focus on sample purification and the quality control of FFPE samples and compare different automated purification procedures.
Critical Factors for Successful Real-Time PCR: Multiplex PCRQIAGEN
Multiplex end-point PCR is a powerful tool for genotyping and many other applications. QIAGEN’s multiplex PCR chemistry is optimized for reliable amplification of many different templates with high variability in copy numbers. Thus it enables very quick establishment of a new lab routine and instant success for your multiplex PCR strategy.
There is a set of critical factors which we recommend to be regarded for planning and performing this kind of PCR. These will be discussed in detail in the webinar. Additionally, our multiplex PCR chemistry has recently been gaining increasing popularity among scientists who are utilizing it for their next-generation sequencing workflows.
Quality control lecture CPath master 2014 Ain ShamsAdel Elazab Elged
Basics of quality management or assurance program detailing values of internal quality control material analysis and interpretation and external quality control or proficiency testing programs in medical laboratories
RIQAS is the largest international External Quality Assessment (EQA)/ Proficiency Testing (PT) scheme, there are currently more than 45,000 participants in 133 countries.
Biological variation as an uncertainty componentGH Yeoh
To assist the clinical interpretation of a test result, there is a necessity to have an additional non-analytical component in the overall estimation of UM, namely the biological variation.
Real-Time quantitative PCR (qPCR) is a mainstream method that is used in research and diagnostic applications for quantification of gene expression. IDT has developed a robust and affordable qPCR master mix for use with probe-based qPCR in single and multiplex assays. In this presentation, we explore a variety of applications of PrimeTime® Gene Expression Master Mix. We cover the use of PrimeTime master mix with probe based assays from IDT. We also look at the use of PrimeTime master mix in multiplex applications without the loss of sensitivity that is commonly observed. Finally, we demonstrate the unmatched stability of PrimeTime master mix under ambient temperatures, saving your research money and minimizing on shipping delays.
Introduction to Real Time PCR (Q-PCR/qPCR/qrt-PCR): qPCR Technology Webinar S...QIAGEN
This slidedeck introduces the concepts of real-time PCR and how to conduct a real-time PCR assay. The topics that are covered include an overview of real-time PCR chemistries, protocols, quantification methods, real-time PCR applications and factors for success.
A guide to lateral flow immunoassay developmentExpedeon
This latest presentation on lateral flow immunoassay development will provide a general overview of lateral flow assays, take you through the components of a typical lateral flow test strip, and will provide you with detail on the different detection methods which are employed. We will also describe how our products and custom services can greatly simplify the development of your lateral flow assay.
To find out more about Innova Biosciences' lateral flow assay development services visit our website:
https://www.innovabiosciences.com/b2b/lateral-flow-assay-development-service.html
ROLE OF IMMUNOASSAY FOR DETECTION OF NARCOTIC DRUG.pptxTereena1
This prestention contains details of different techniques of immunoassays and its details and their use in detection of narcotic drugs such as opiuim and cannabis
This slide show forms part of the Introduction to Flow Cytometry seminar help by The Garvan MLC Flow Cytometry Facility. The Garvan MLC Flow Cytometry Facility is part of the Garvan Institute of Medical Research and is located in Sydney NSW.
This presentation explains about the principle and procedure involved in elisa method of immunoassay, development o f elisa , application advantages and disadvantages of elisa
Nucleic Acid Quantification from FFPE Samples – Are You Doing it Right?QIAGEN
Formalin-fixation and paraffin-embedding is a standard method for long-term preservation of tissue biopsies and these stored samples are a valuable tool for studying diseases such as cancer, especially when they are histologically and pathologically well characterized, and follow-up clinical data is available. The quality of nucleic acids extracted from FFPE samples is influenced by a number of factors, including how the samples were handled before, during and after fixation and embedding. Moreover, there are several difficulties when purifying nucleic acids from FFPE samples as the chemicals and temperature used during the process can degrade the DNA.
In this webinar, we will discuss the variability in quantity and purity of DNA purified from FFPE material. We will show data from different quantification and quality control methods, demonstrate the impact of inaccurate quantification on downstream results and discuss how to overcome these challenges.
DNA sequence analysis and genotyping of biological samples using innovative instrumentation, such as next-generation sequencing (NGS) platforms, is often limited by the small amount of sample available. The REPLI-g Single Cell Kit is specially designed to uniformly amplify genomic DNA from single cells (1 to <1000 bacterial or tumor cells) or purified genomic DNA with complete genome coverage. Additional protocols are also available for use with fresh or dried blood or fresh or frozen tissue. Dedicated buffers and reagents undergo a unique, controlled decontamination procedure to avoid amplification of contaminating DNA, ensuring highly reliable results every time. Accurate amplification of genomes with negligible sequence bias and no genomic drop-outs is achieved with innovative Multiple Displacement Amplification (MDA) technology. In contrast to PCR-based WGA technologies, high fidelity rates are increased up to 1000-fold, avoiding costly false positive or negative results.
RIQAS is the largest international External Quality Assessment (EQA)/ Proficiency Testing (PT) scheme, there are currently more than 45,000 participants in 133 countries.
Biological variation as an uncertainty componentGH Yeoh
To assist the clinical interpretation of a test result, there is a necessity to have an additional non-analytical component in the overall estimation of UM, namely the biological variation.
Real-Time quantitative PCR (qPCR) is a mainstream method that is used in research and diagnostic applications for quantification of gene expression. IDT has developed a robust and affordable qPCR master mix for use with probe-based qPCR in single and multiplex assays. In this presentation, we explore a variety of applications of PrimeTime® Gene Expression Master Mix. We cover the use of PrimeTime master mix with probe based assays from IDT. We also look at the use of PrimeTime master mix in multiplex applications without the loss of sensitivity that is commonly observed. Finally, we demonstrate the unmatched stability of PrimeTime master mix under ambient temperatures, saving your research money and minimizing on shipping delays.
Introduction to Real Time PCR (Q-PCR/qPCR/qrt-PCR): qPCR Technology Webinar S...QIAGEN
This slidedeck introduces the concepts of real-time PCR and how to conduct a real-time PCR assay. The topics that are covered include an overview of real-time PCR chemistries, protocols, quantification methods, real-time PCR applications and factors for success.
A guide to lateral flow immunoassay developmentExpedeon
This latest presentation on lateral flow immunoassay development will provide a general overview of lateral flow assays, take you through the components of a typical lateral flow test strip, and will provide you with detail on the different detection methods which are employed. We will also describe how our products and custom services can greatly simplify the development of your lateral flow assay.
To find out more about Innova Biosciences' lateral flow assay development services visit our website:
https://www.innovabiosciences.com/b2b/lateral-flow-assay-development-service.html
ROLE OF IMMUNOASSAY FOR DETECTION OF NARCOTIC DRUG.pptxTereena1
This prestention contains details of different techniques of immunoassays and its details and their use in detection of narcotic drugs such as opiuim and cannabis
This slide show forms part of the Introduction to Flow Cytometry seminar help by The Garvan MLC Flow Cytometry Facility. The Garvan MLC Flow Cytometry Facility is part of the Garvan Institute of Medical Research and is located in Sydney NSW.
This presentation explains about the principle and procedure involved in elisa method of immunoassay, development o f elisa , application advantages and disadvantages of elisa
Nucleic Acid Quantification from FFPE Samples – Are You Doing it Right?QIAGEN
Formalin-fixation and paraffin-embedding is a standard method for long-term preservation of tissue biopsies and these stored samples are a valuable tool for studying diseases such as cancer, especially when they are histologically and pathologically well characterized, and follow-up clinical data is available. The quality of nucleic acids extracted from FFPE samples is influenced by a number of factors, including how the samples were handled before, during and after fixation and embedding. Moreover, there are several difficulties when purifying nucleic acids from FFPE samples as the chemicals and temperature used during the process can degrade the DNA.
In this webinar, we will discuss the variability in quantity and purity of DNA purified from FFPE material. We will show data from different quantification and quality control methods, demonstrate the impact of inaccurate quantification on downstream results and discuss how to overcome these challenges.
DNA sequence analysis and genotyping of biological samples using innovative instrumentation, such as next-generation sequencing (NGS) platforms, is often limited by the small amount of sample available. The REPLI-g Single Cell Kit is specially designed to uniformly amplify genomic DNA from single cells (1 to <1000 bacterial or tumor cells) or purified genomic DNA with complete genome coverage. Additional protocols are also available for use with fresh or dried blood or fresh or frozen tissue. Dedicated buffers and reagents undergo a unique, controlled decontamination procedure to avoid amplification of contaminating DNA, ensuring highly reliable results every time. Accurate amplification of genomes with negligible sequence bias and no genomic drop-outs is achieved with innovative Multiple Displacement Amplification (MDA) technology. In contrast to PCR-based WGA technologies, high fidelity rates are increased up to 1000-fold, avoiding costly false positive or negative results.
Simultaneous Isolation of RNA & DNA from one FFPE SampleQIAGEN
Worldwide, there are millions of tissue samples archived in tissue biobanks and biorepositories. These samples are extremely valuable for pharmacological and biomedical research and companion diagnostics, due to the linkage to patient history. The vast majority of archived tissue samples are formalin-fixed and paraffin-embedded (FFPE), since formalin is the standard fixative for tissue samples.
FFPE blocks serve as an excellent source for histomorphology studies, but their use in molecular studies is challenging, due to crosslinking and fragmentation caused by fixation, processing, embedding, and storage conditions. For reliable comparison of genomic and transcriptomic data from heterogeneous samples and to spare sample material, purification of DNA and RNA from
the same sample is essential. This is particularly important when working with tumorous tissues, which contain a heterogeneous distribution of healthy and malignant cells.
Targeted RNAseq for Gene Expression Using Unique Molecular Indexes (UMIs): In...QIAGEN
Traditional RNA sequencing (RNA-Seq) is a powerful tool for expression profiling, but is hindered by PCR amplification bias and inaccuracy at low expressing genes. QIAseq RNA is a flexible and precise tool developed for mitigating these complications, allowing digital gene expression analysis. This in-depth webinar will cover sample requirements, experimental design, NGS platform-specific challenges and workflow for gene enrichment, library prep and sequencing. The applications of QIASeq RNA Panels in cancer research, stem cell differentiation and elucidating the effects small molecules on signaling pathways will be highlighted.
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
Comparing Mutation Detection Sensitivity from Matched FFPE Tissue and Liquid ...Thermo Fisher Scientific
Cancer researchers are avidly working to enable circulating cell free DNA (cfDNA) profiling as a new more sensitive tool to detect and screen for the presence of solid tumors before detection through clinical methods. Despite the high level of interest in cfDNA, researchers still have reservations until enough data has demonstrated complementarity between methodologies. In this study, we examined the data quality and concordance of mutations called for a small number of matched formalin fixed paraffin embedded (FFPE) tissue and plasma samples.
Polymerase chain reaction (abbreviated PCR) is a laboratory technique for rapidly producing (amplifying) millions to billions of copies of a specific segment of DNA, which can then be studied in greater detail. PCR involves using short synthetic DNA fragments called primers to select a segment of the genome to be amplified, and then multiple rounds of DNA synthesis to amplify that segment. This slides introduces pcr importances ,uses and steps of pcr.
Cancer therapies that target specific pathways can be more effective than established, nonspecific chemotherapy and radiation treatments, and may prevent side effects on healthy tissues. Such targeted therapies can only be applied after underlying gene mutations have been identified. However, detecting low frequency variants from clinically relevant samples poses significant challenges. Specimens are routinely formalin-fixed and paraffin-embedded (FFPE) for histology, which can decrease the efficiency of NGS library preparation. In this presentation, we discuss approaches for extraction of DNA from FFPE samples, and recommend quality control assays to guide parameter selection for library construction and sequencing depth.
High-throughput processing to maximize genomic analysis through simultaneous ...Thermo Fisher Scientific
As personalized cancer care evolves, the patient’s nucleic acid becomes ever so important to provide valuable information regarding their genetic makeup and disease state. Common sample types for these analyses include biopsies, which can be very limited in material making the downstream measurement of more than one analyte rather difficult. Obtaining another biopsy, using a different section or splitting the sample can be problematic because of tumor heterogeneity. Even adjacent areas of the same tumor tissue can result in different RNA/DNA profiles so the ability to isolate multiple analytes from the same sample offer a number of benefits, which include preserving samples and data consistency eliminating any sample to sample variation. As more tests are developed to simultaneously monitor genetic alterations, there is a strong need to efficiently isolate both DNA and RNA from the same starting sample in a format compatible with high-throughput processing.
In this slides the topic that which is discussed is "How PCR is involved in identification of Genotype"
I hope this will Help you in your presentation work.
"PCR can be used in identification of genotype."
Step by Step, from Liquid Biopsy to a Genomic Biomarker: Liquid Biopsy Series...QIAGEN
Liquid biopsies enable us to monitor the evolution of genetic aberrations in primary tumors as they shed the tumor cells into the circulation. The limitation is the ability to detect these low frequency genetic aberrations in a consistent manner to understand short- and long-term implications and how this information will be used in the clinic. This slidedeck will cover the challenges and solutions associated with multiple steps as one starts with liquid biopsy and move towards finding a new biomarker.
Similar to Challenges of FFPE Sample Materials – Where Does Variation in Quantity of Purified DNA Come From? (20)
Using methylation patterns to determine origin of biological material and ageQIAGEN
In this QIAGEN sponsored webinar, our guest speakers from the San Francisco Police Department (SFPD) Crime Lab and Florida International University (FIU) discuss their research on the potential of epigenetic methylation as a procedure for body fluid identification and age estimation from DNA left at crime scenes. Several approaches have been studied, including an analysis of methyl array data and an initial validation of procedures such as pyrosequencing and real-time PCR. The presentation focuses on a number of tissue-specific epigenetic markers for body fluid and age determination with a promise of future integration of these markers into the forensic lab due to the simplicity of analysis and the ease of application.
Learn more about the Pyrosequencing technology and our solutions at
https://www.qiagen.com/resources/technologies/pyrosequencing-resource-center/
Take lung cancer research to a new molecular dimensionQIAGEN
Circulating Tumor Cells (CTCs) can provide researchers with important new discoveries on the mechanism of cancer. Find out more about the latest technology that provides researchers the necessary tools to conduct CTC research in lung cancer.
Circulating Tumor Cells (CTCs) can provide researchers with important new discoveries on the mechanism of cancer. Find out more about the latest technology that provides researchers the necessary tools to conduct CTC research in AR-V7 related prostate cancer.
Learn about the power of LNA (Locked Nucleic Acid) technology and QIAGEN's LNA enhanced product portfolio for RNA and DNA research. Download the slide deck!
Take your RNA research to the next level with QIAGEN LNA tools!QIAGEN
Download the flyer!
Experience truly exceptional RNA research with QIAGEN's next-generation, LNA®-enhanced tools. LNA (Locked Nucleic Acid) oligos bind with much higher affinity and specificity to RNA targets than standard DNA and RNA oligos – This enables specific and sensitive detection of small RNAs and discrimination between highly similar
sequences.
An Approach to De-convolution of Mixtures in Touch DNA Samples. Download now!QIAGEN
7th QIAGEN Investigator Forum - Lisbon, March 8, 2018 . An Approach to De-convolution of Mixtures in Touch DNA Samples. Presenter: Lisa Dierig, Institute of Legal Medicine, Ulm
Assessment of Y chromosome degradation level using the Investigator® Quantipl...QIAGEN
Assessment of Y chromosome degradation level using the Investigator® Quantiplex® Pro RGQ Kit, presented by Dr. Tomasz Kupiec, Head of the Forensic Genetics Section, Institute of Forensic Research, Krakow, Poland on June 14, 2018.
ICMP MPS SNP Panel for Missing Persons - Michelle Peck et al.QIAGEN
Optimization and Performance of a Very Large MGS SNP Panel for Missing Persons, by Michelle Peck et al., International Commission on Mission Persons. Presented May 3, 2018, at the QIAGEN Investigator Forum, San Antonio, TX.
Exploring the Temperate Leaf Microbiome: From Natural Forests to Controlled E...QIAGEN
The aerial surfaces of plants, the phyllosphere, harbors a diverse community of microorganisms. The increasing awareness of the potential roles of phyllosphere microbial communities calls for a greater understanding of their structure and dynamics in natural and urban ecosystems. To do so, we characterized the community structure and assembly dynamics of leaf bacterial communities in natural temperate forests of Quebec by comparing the relative influence of host species identity, site, and time on phyllosphere bacterial community structure. Second, we tested the value of characterizing a tree’s complete phyllosphere microbial community through a single sample by measuring the intra-individual, inter-individual and interspecific variation in leaf bacterial communities. Third, we quantified the relationships among phyllosphere bacterial diversity, plant species richness, plant functional diversity and identity, and plant community productivity in a biodiversity-ecosystem function experiment with trees. Finally, we compared tree leaf bacterial communities in natural and urban environments, as well as along a gradient of increasing anthropogenic pressures. The work presented here thus offers an original assessment of the dynamics at play in the tree phyllosphere.
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.
The Microbiome of Research Animals : Implications for Reproducibility, Transl...QIAGEN
The human gut microbiota (GM) has emerged as a key factor in susceptibility to, as well as a potential biomarker of, several diseases and conditions. Similarly, researchers now appreciate that the GM of laboratory animals could affect the reproducibility and translatability of many disease models, including a complete loss of phenotype. While associations between characteristics of the GM and differential disease model phenotypes are of concern, they can also be viewed as sources of discovery related to disease pathogenesis. As such, there is considerable interest in factors that inadvertently influence the composition of the GM and methods of manipulating the GM prospectively to investigate such associations and standardize or optimize disease models. The webinar will present data on variables capable of influencing the GM of laboratory rodents citing several examples and animal models, considerations related to manipulation of the GM in mice and rats, and recent data supporting the use of “dirty” mice in biomedical research.
Building a large-scale missing persons ID SNP panel - Download the studyQIAGEN
In this webinar, we will take a look at a large-scale SNP-based forensic identification panel for DNA analysis with massively parallel sequencing (MPS). The panel was specifically designed for the challenges of identifying missing persons; where DNA is frequently highly degraded, and relationship tests may involve reference samples from across several generations and in a deficient pedigree.
Rapid DNA isolation from diverse plant material for use in Next Generation Se...QIAGEN
Isolation of DNA from plant material is often a tedious process which involves significant hands on time and leads to varying results due to the diverse nature of the material. Different parts of the plants as well as the plants themselves differ in both consistency of material and presence of inhibitory substances, making dependable isolation of DNA difficult.
Here, we developed a method for the efficient extraction of DNA from different plant types, including strawberry leaf, pine needle, grape leaf, and cotton and coffee seeds (workflow at right). A novel bead beating method and lysis chemistry led to more efficient sample lysis with minimal hands-on time and significantly increased DNA yield compared to conventional methods. Through the use of multiple technologies to improve removal of secondary metabolites, such as polyphenols, complex polysaccharides, alkaloids and tannins that may inhibit downstream applications, the isolated DNA was of high quality and purity.
The resulting DNA is suitable for immediate use in downstream reactions, including PCR, qPCR and Next Generation Sequencing based applications. Using this method we were further able to design a workflow that included DNA isolation, library preparation and bioinformatics analyses for the efficient detection of plant pathogens isolated from infected samples. With this, our protocol is a substantial improvement within workflows used for plant microbiome and plant pathology studies as well as in plant breeding and engineering.
Rapid extraction of high yield, high quality DNA from tissue samples - Downlo...QIAGEN
Genetic and genomic analysis from tissue samples requires the extraction of high quality DNA. Mechanical disruption methods such as bead milling provide high yield from tissue samples, but cause damage to the nucleic acids. Purely enzymatic methods such as proteinase K digestion can extract nucleic acid without damage, but require long incubation times, often proceeding overnight, and without approaching the yields achieved by mechanical disruption techniques. Thus a method is needed which can provide a rapid extraction of high yield, high quality DNA from tissue samples. See the new method.
Practical hints and new solutions for successful real-time PCR studies QIAGEN
Part 1: Practical hints and new solutions for successful real-time PCR studies
In this webinar we will cover the following topics which are critical steps for efficient and precise gene expression studies using real-time PCR technology:
- Effect of RNA integrity on real-time PCR results – tips to achieve a true RNA profiling suitable for real-time PCR studies
- Improved methods for cDNA synthesis, optimized for real-time PCR
- Real-time PCR analysis
o Real-time PCR essentials and background information on different quantification strategies
o SYBR Green real-time PCR – factors influencing specificity
o Introduction to probe technology
o New, fast and efficient real-time PCR solutions
Part 2: Critical Factors for Successful Multiplex Real-Time PCR
Multiplex real-time PCR is a powerful tool for gene expression analysis, viral load monitoring, genotyping, and many other applications. The ability to amplify and detect several genomic DNA, cDNA, or RNA targets in the same reaction offers many benefits:
• Conservation of precious samples – more quantification data per sample
• Increased throughput – more targets analyzed per run on a cycler
• Reliable results – no well-to-well variability due to co-amplification of internal control
• Reduced costs – save time and reagents
The QuantiFast Multiplex PCR and RT-PCR kits are optimized for reliable amplification of many different templates despite a high variability in abundance. Thus they enable successful amplification of multiple targets on the first attempt without optimization.
This webinar explains the principles of the QIAGEN multiplex technologies and shows data demonstrating the exceptional multiplex real-time PCR performance of the QuantiFast Multiplex kits.
Overcome the challenges of Nucleic acid isolation from PCR inhibitor-rich mic...QIAGEN
This presentation will focus on nucleic acid extraction tools developed by QIAGEN that facilitate accurate non-biased community analysis and eliminate common amplification problems via the depletion of endogenous polymerase inhibitors using our patented Inhibitor Removal Technology.
RotorGene Q A Rapid, Automatable real-time PCR Instrument for Genotyping and...QIAGEN
QIAGEN has developed a selection of robust, novel chemistries to prevent PCR crosstalk. We can successfully measure target abundance and fold change in real-time assays, and perform sub-genotyping using a fast, high-throughput and powerful High-Resolution Melting (HRM) statistical analysis program. In this presentation, we will demonstrate these features and benefits with examples.
Reproducibility, Quality Control and Importance of AutomationQIAGEN
In this webinar, we will introduce you to the key sample quality parameters, discuss their respective impact on downstream applications and how to monitor them, and present the advantages of automating quality control along complex workflows.
Automated Nucleic Acid Purification from Diverse Sample types using dedicated...QIAGEN
This webinar will focus on the automation of QIAGEN’s new line of DNA and RNA sample prep kits for the microbiome. We will show how automation on the QIAcube enables efficient and reliable use of these samples for sensitive downstream applications such as qPCR and NGS. In addition, you will learn how to successfully use the CLC Microbial Genomics Module for metagenome sequencing and identification of microbial composition and diversity.
Dna Methylation Analysis in a Single Day - Download the SlidesQIAGEN
This webinar introduces the new PyroMark Q48 Autoprep system. Combined with the latest EpiTect Fast bisulfite conversion technology, the new PyroMark Q48 Autoprep can now provide highly automated methylation analysis in a single day.
CHAPTER 1 SEMESTER V - ROLE OF PEADIATRIC NURSE.pdfSachin Sharma
Pediatric nurses play a vital role in the health and well-being of children. Their responsibilities are wide-ranging, and their objectives can be categorized into several key areas:
1. Direct Patient Care:
Objective: Provide comprehensive and compassionate care to infants, children, and adolescents in various healthcare settings (hospitals, clinics, etc.).
This includes tasks like:
Monitoring vital signs and physical condition.
Administering medications and treatments.
Performing procedures as directed by doctors.
Assisting with daily living activities (bathing, feeding).
Providing emotional support and pain management.
2. Health Promotion and Education:
Objective: Promote healthy behaviors and educate children, families, and communities about preventive healthcare.
This includes tasks like:
Administering vaccinations.
Providing education on nutrition, hygiene, and development.
Offering breastfeeding and childbirth support.
Counseling families on safety and injury prevention.
3. Collaboration and Advocacy:
Objective: Collaborate effectively with doctors, social workers, therapists, and other healthcare professionals to ensure coordinated care for children.
Objective: Advocate for the rights and best interests of their patients, especially when children cannot speak for themselves.
This includes tasks like:
Communicating effectively with healthcare teams.
Identifying and addressing potential risks to child welfare.
Educating families about their child's condition and treatment options.
4. Professional Development and Research:
Objective: Stay up-to-date on the latest advancements in pediatric healthcare through continuing education and research.
Objective: Contribute to improving the quality of care for children by participating in research initiatives.
This includes tasks like:
Attending workshops and conferences on pediatric nursing.
Participating in clinical trials related to child health.
Implementing evidence-based practices into their daily routines.
By fulfilling these objectives, pediatric nurses play a crucial role in ensuring the optimal health and well-being of children throughout all stages of their development.
For those battling kidney disease and exploring treatment options, understanding when to consider a kidney transplant is crucial. This guide aims to provide valuable insights into the circumstances under which a kidney transplant at the renowned Hiranandani Hospital may be the most appropriate course of action. By addressing the key indicators and factors involved, we hope to empower patients and their families to make informed decisions about their kidney care journey.
Navigating Challenges: Mental Health, Legislation, and the Prison System in B...Guillermo Rivera
This conference will delve into the intricate intersections between mental health, legal frameworks, and the prison system in Bolivia. It aims to provide a comprehensive overview of the current challenges faced by mental health professionals working within the legislative and correctional landscapes. Topics of discussion will include the prevalence and impact of mental health issues among the incarcerated population, the effectiveness of existing mental health policies and legislation, and potential reforms to enhance the mental health support system within prisons.
CHAPTER 1 SEMESTER V PREVENTIVE-PEDIATRICS.pdfSachin Sharma
This content provides an overview of preventive pediatrics. It defines preventive pediatrics as preventing disease and promoting children's physical, mental, and social well-being to achieve positive health. It discusses antenatal, postnatal, and social preventive pediatrics. It also covers various child health programs like immunization, breastfeeding, ICDS, and the roles of organizations like WHO, UNICEF, and nurses in preventive pediatrics.
LGBTQ+ Adults: Unique Opportunities and Inclusive Approaches to CareVITASAuthor
This webinar helps clinicians understand the unique healthcare needs of the LGBTQ+ community, primarily in relation to end-of-life care. Topics include social and cultural background and challenges, healthcare disparities, advanced care planning, and strategies for reaching the community and improving quality of care.
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India Diagnostic Labs Market: Dynamics, Key Players, and Industry Projections...Kumar Satyam
According to the TechSci Research report titled “India Diagnostic Labs Market Industry Size, Share, Trends, Competition, Opportunity, and Forecast, 2019-2029,” the India Diagnostic Labs Market was valued at USD 16,471.21 million in 2023 and is projected to grow at an impressive compound annual growth rate (CAGR) of 11.55% through 2029. This significant growth can be attributed to various factors, including collaborations and partnerships among leading companies, the expansion of diagnostic chains, and increasing accessibility to diagnostic services across the country. This comprehensive report delves into the market dynamics, recent trends, drivers, competitive landscape, and benefits of the research report, providing a detailed analysis of the India Diagnostic Labs Market.
Collaborations and Partnerships
Collaborations and partnerships among leading companies play a pivotal role in driving the growth of the India Diagnostic Labs Market. These strategic alliances allow companies to merge their expertise, strengthen their market positions, and offer innovative solutions. By combining resources, companies can enhance their research and development capabilities, expand their product portfolios, and improve their distribution networks. These collaborations also facilitate the sharing of technological advancements and best practices, contributing to the overall growth of the market.
Expansion of Diagnostic Chains
The expansion of diagnostic chains is a driving force behind the growing demand for diagnostic lab services. Diagnostic chains often establish multiple laboratories and diagnostic centers in various cities and regions, including urban and rural areas. This expanded network makes diagnostic services more accessible to a larger portion of the population, addressing healthcare disparities and reaching underserved populations. The presence of diagnostic chain facilities in multiple locations within a city or region provides convenience for patients, reducing travel time and effort. A broader network of labs often leads to reduced waiting times for appointments and sample collection, ensuring that patients receive timely and efficient diagnostic services.
Rising Prevalence of Chronic Diseases
The increasing prevalence of chronic diseases is a significant driver for the demand for diagnostic lab services. Chronic conditions such as diabetes, cardiovascular diseases, and cancer require regular monitoring and diagnostic testing for effective management. The rise in chronic diseases necessitates the use of advanced diagnostic tools and technologies, driving the growth of the diagnostic labs market. Additionally, early diagnosis and timely intervention are crucial for managing chronic diseases, further boosting the demand for diagnostic lab services.
How many patients does case series should have In comparison to case reports.pdfpubrica101
Pubrica’s team of researchers and writers create scientific and medical research articles, which may be important resources for authors and practitioners. Pubrica medical writers assist you in creating and revising the introduction by alerting the reader to gaps in the chosen study subject. Our professionals understand the order in which the hypothesis topic is followed by the broad subject, the issue, and the backdrop.
https://pubrica.com/academy/case-study-or-series/how-many-patients-does-case-series-should-have-in-comparison-to-case-reports/
Challenges of FFPE Sample Materials – Where Does Variation in Quantity of Purified DNA Come From?
1. Sample to Insight
The challenges of FFPE sample materials – where does
variation in quantity of purified DNA come from?
Dr. Carola Schade, QIAGEN GmbH
3. Sample to Insight
Some facts
3
Trends
FFPE has become a standard method for long-term preservation of tissue biopsies
Large number of unprocessed FFPE samples are archived in tissue banks and
biorepositories
Samples are highly valuable, especially when they are well characterized
Needs
Maximum recovery from precious, small FFPE samples
DNA must be suitable for all types of applications, including NGS
Removal of co-purified RNA (i.e., for DNA sequencing)
Differentiation between artificial and true mutation
Sometimes there is no other choice than FFPE
4. Sample to Insight
Important considerations
4
Sample handling
Time between excision and fixation
Changes in RNA transcript profile and proteins
Formalin fixation
pH of solution, composition of fixative
Thickness of tissue specimen
Duration of fixation process, volume of formalin
solution
Acid-mediated hydrolytic nucleic acid fragmentation
Cytosine deamination
Crosslinking of biomolecules
Sample
handling
Deparaffin-
ization
Embedding StorageFixation Purification
Molecular
analysis
Preparation and archiving Retrieval and analysis
Embedding
Full dehydration
Melting characteristics of paraffin used
Residual water can lead to proteolysis
Sample degradation when using high-melting
temperature paraffin
Storage
Storage temperature
Sample degradation
Challenges when working with FFPE samples
5. Sample to Insight
QIAamp DNA FFPE Tissue Kit
5
Kit specifications
Silica membrane based
Up to 8 sections, each with a thickness of up to
10 µm and a surface area of up to 250 mm2
Purification of genomic DNA & mitochondrial DNA
Elution volume 20–100µl
QIAcube protocol available
Paraffin removal and sample lysis
No need for overnight incubation
Paraffin is dissolved in xylene and removed
Sample lysis under denaturing conditions with
proteinase K (1 h, 56°C)
Incubation at 90°C to reverse formalin crosslinking
Optional RNase treatment step
7. Sample to Insight
Multiple studies investigating variation in FFPE sample processing
7
What impacts downstream results most?
8. Sample to Insight
Some conflicting messages
8
Tissue type(s) not specified
RNase digest Yes/No unclear
Relative yields vs. absolute yields
Systematic deviations in quantification
dependent on method used? “No method
highly superior
to others...”
... it is particularly important to
choose the most reliable and
constant DNA extraction system,
especially when using small
biopsies and low elution volumes...
“ ...variation in pre-PCR steps is
prevalent...”
...all common DNA quantification
techniques can be used for
downstream applications...
“DNA
quantitation may
also impact PCR
efficiency...”
10. Sample to Insight
Which factor has the highest impact on DNA quantity measured?
10
Study design Samples
5 different rat tissue types
2–3 different blocks
5-6x 3 sections, 10 µm thickness
Assess variability caused by the samples
themselves
FFPE Samples
Nanodrop QIAxpert Qubit
In total 6000 data points!
QIAcube Manual
w/ RNase digest w/o RNase digest
DNA purification
Automated using the QIAcube or manual
processing
QIAamp DNA FFPE Kit
With and without RNase digest
Assess variability introduced by operator
(manual vs. automated)
DNA quantification
Using three different methods,
5 replicates/sample
Assess variability caused by downstream
quantification method
11. Sample to Insight
DNA quantification technologies
11
Nanodrop QIAxpert Qubit
Technology
UV/VIS absorbance
reading
UV/VIS absorbance
reading
Fluorescence-based assay
LOD 2 ng/µl (dsDNA) 1.5 ng/µl 10 pg/µl (assay dependent)
Sample volume 1 µl 2 µl 1–20 µl
Measurements needed
for 16 samples
16 1 16
Drop-and-clean actions
required
Yes No No
Reported values
A260 Yes Yes
A280 Yes Yes
A260/280 Yes Yes
A260/230 Yes Yes
Discriminate between
molecules of interest
No Yes (Yes)
12. Sample to Insight
DNA – measurement accuracy
14
173 ng/µl reference DNA (Promega ultra pure gDNA )
All technologies with slight
underquantification
Low %CV value for QIAxpert
(Nanodrop with lower no. of
samples being measured)
Qubit with high mean variation
How was the initial
concentration
determined by
Promega?
Accuracy
Reference DNA (Promega ultra pure gDNA ) was used at a concentration of 173 ng/µL and 5 ng/µl (dilution from original
solution in H2O). A total of 63 replicates were measured on two different QIAxpert systems, 32 replicates were measured on
a Nanodrop 8000, and 15 replicates were additionally measured using the Qubit system.
Instrument Sample
no.
AVG
[ng/µl]
STD
[ng/µl]
%CV
Nanodrop8000 32 167.6 1.7 0.6
QIAxpert 63 163.9 2.4 0.9
Qubit 15 154.9 11.1 4.6
13. Sample to Insight
DNA – measurement accuracy
15
5 ng/µl reference DNA (Promega ultra pure gDNA )
All technologies with slight
underquantification
Mean variation increases for absorbance
reading
Qubit variance seems to be lower,
however, also smaller sample number
that was measured
How was the initial concentration
determined by Promega?
Dilution of original solution may
influence accuracy of
measurement.
Accuracy
Reference DNA (Promega ultra pure gDNA ) was used at a concentration of 173 ng/µL and 5 ng/µl (dilution from original
solution in H2O). A total of 63 replicates were measured on two different QIAxpert systems, 32 replicates were measured on
a Nanodrop 8000, and 15 replicates were additionally measured using the Qubit system.
14. Sample to Insight
DNA – comparison of linearity using different systems
14
QIAxpert Nanodrop Qubit
Linearity
Calf thymus DNA (Life Technolgies) was purified using QIAamp chemistry on the QIAcube, and a serial dilution of the final eluate (137 ng/µl) was generated,
representing 120 ng/µl, 100 ng/µl, 80 ng/µl, 60 ng/µl, 50 ng/µl, 40 ng/µl, and 30 ng/µl. A total of 5 replicates of each dilution were subsequently measured using the
QIAxpert system, a Nanodrop 8000, and the Qubit. Data shown for the QIAxpert reflects total NA measured with the dsDNA QIAamp app.
Nanodrop with a systematic overquantification
Qubit with a systematic underquantification
15. Sample to Insight
Which factor has the highest impact on DNA quantity measured?
15
Study design Samples
5 different rat tissue types
2-3 different blocks
5-6 x 3 sections, 10µm thickness
Assess variability caused by samples
themselves
DNA Purification
Automated using the QIAcube or manual
processing
QIAamp DNA FFPE Kit
w/ and w/o RNase digest
Assess variability introduced by operator
(manual vs. automated)
DNA Quantification
Using three different methods,
5 replicates/sample
Assess variability caused by downstream
quantification method
FFPE Samples
QIAcube Manual
Nanodrop QIAxpert Qubit
In total 6000 data points !
w/ RNase digest w/o RNase digest
16. Sample to Insight
Comparison of different quantification systems
16
Concentration variability of purified FFPE samples
What are the factors contributing to this variability when doing absorbance
readings?
QIAxpertNanodrop Qubit
Nanodrop QIAxpert Qubit
Sample no. 1500 3000 1500
AVG [ng/µl] 66.4 49.4 14.5
STD [ng/µl] 98.8 67.7 10.3
A huge variability with all UV/VIS-based
systems
Nanodrop shows the highest variance
Qubit with the lowest variance
17. Sample to Insight
Does RNase treatment have an influence?
17
What about automated vs. manual processing?
QIAxpertNanodrop Qubit
Nanodrop QIAxpert Qubit
RNase
digest
w/o w/ w/o w/ w/o w/
AVG [ng/µl] 103.2 29.6 82.9 15.7 16.1 13.0
STD [ng/µl] 125.9 31.4 82.3 11.7 11.2 9.1
A huge influence of variation is related to RNA
QIAxpert and Qubit show similar low variance on
RNase-treated samples
Nanodrop shows high variance on RNase-
treated and untreated samples
Concentration variability of RNase-treated FFPE samples
with RNase digest
without RNase digest
18. Sample to Insight
Does the kind of purification have an influence?
18
Concentration variability with kind of purification
What role does the tissue type play?
QIAxpertNanodrop Qubit
Kind of purification
Automated Manual
Kind of purification
Automated Manual
Kind of purification
Automated Manual
Higher level of standardization applying
automated sample purification
with RNase digest
without RNase digest
19. Sample to Insight
Comparison of different FFPE tissue types
19
Concentration variability among different FFPE tissue samples
Rat colon Rat heart Rat kidney Rat liver Rat muscle
Min. conc. [ng/µl] 0 3.6 2.7 0.8 0.3
Max. conc. [ng/µl] 193.3 117.9 252.9 810.3 60.3
STD [ng/µl] 35.4 21.1 71.4 125.0 13.5
Different FFPE tissue material leads to different yield of nucleic acids
Liver tissue is most challenging because of higher portion of RNA
20. Sample to Insight
20
Comparison of different FFPE tissue types
Concentration variability among different FFPE tissue types
Variability of nucleic acid
concentration determination is
related to different amounts of
RNA
QIAxpert and Qubit show
similar low variance on RNase-
treated samples
Nanodrop shows high variance
on RNase-treated and
untreated samples
Main contriubtion to variability related to RNA amounts
21. Sample to Insight
What is the contribution to the overall variability by the block/section?
Comparison of different FFPE tissue types
21
Less variability when purification is automated
Variability of nucleic acid
concentration determination is
related to different amounts of
RNA
Higher level of standardization
applying automated sample
purification
22. Sample to Insight
Contribution to variability in quantification by the block/section
22
Higher differences in yields due to the quantification method chosen
rather than block or section
23. Sample to Insight
Contribution to variability in quantification by the block/section
23
Higher differences in yields due to the quantification method chosen
rather than block or section
24. Sample to Insight
What has the highest influence on quantification of nucleic acids purified
from FFPE samples?
24
%Contribution
Quantification technology 0.2816
Purification method 0.2569
Tissue type 0.2288
RNase digest 0.1424
FFPE block 0.0542
FFPE section 0.0362
The chosen quantification technology matters most!
26. Sample to Insight
Summary
26
FFPE tissue samples present a number of challenges
If you really want to be sure that the genomic DNA you quantify represents what
is in your sample:
Choose your quantification technology carefully
Automate your sample prep (i.e., using a QIAcube)
Apply a RNase digestion step
Be aware of systematic differences between technologies when quantifying
nucleic acids
QIAxpert system offers reliable quantification of FFPE samples
QIAGEN provides a number of solutions – from Sample to Insight –
supporting your research efforts using FFPE samples
27. Sample to Insight
Q&A session
27
Thank you for your attention!
Questions?
For up-to-date licensing information and product-specific disclaimers for QIAGEN products,
see the respective QIAGEN kit handbook or user manual. QIAGEN kit handbooks and user
manuals are available at www.qiagen.com or can be requested from QIAGEN Technical
Services or your local distributor.