RT2 Profiler PCR Arrays are a real-time PCR technology that allows researchers to study gene expression patterns across biological pathways and processes. The arrays contain pre-designed primer assays for 84 relevant genes as well as controls on a single plate in a 96-well format. The gene content of the arrays is selected based on biological relevance and published associations with relevant pathways. The primer assays on the arrays undergo extensive validation for sensitivity, specificity, reproducibility, and amplification efficiency. The PCR Array system also includes optimized components for RNA isolation, cDNA synthesis, and real-time PCR to provide a complete validated workflow for gene expression analysis from sample to results.
PCR Array Data Analysis Tutorial: qPCR Technology Webinar Series Part 3QIAGEN
Using actual PCR Array data, this slidedeck presents an easy-to-use and free web-based data analysis tool to calculate fold-differences in gene expression from your raw real-time PCR threshold cycles. Learn how you can look at your results in different formats, including heat map, scatter, volcano, clustergram and multigroup plot.
This slidedeck presents a simple and accurate real-time PCR system for relevant biological pathway- and disease-focused mRNA and long noncoding RNA (lncRNA) expression profiling. Learn about the stringent performance built into the technology to ensure its sensitivity, specificity, reproducibility and reliability. Application examples are also presented.
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!
New Progress in Pyrosequencing for DNA MethylationQIAGEN
Pyrosequencing is a highly flexible technology that lets you rapidly analyze short- to medium-length sequences fast and quantitatively with high accuracy. The real-time, high-resolution sequence output makes the technology highly suitable for applications including complex mutation analysis, microbial identification and DNA methylation quantification.
The main bottleneck in Pyrosequencing has been limited sequence length, which is critical for some applications. Our new technology, software, and chemistry overcome this bottleneck and give sequence reads that are typically twice as long as those from previous PyroMark systems. The new PyroMark Q24 Advanced system also reduces background noise, improving quantification even at sites distant from the sequencing start. The new system is ideal for applications requiring analysis of longer sequences, such as DNA methylation analysis in epigenetic research, frequency determination in mutation analysis, and various de novo sequencing applications.
In this presentation, we will discuss the following applications and technology improvements:
• DNA methylation analysis at single base resolution at CpG and CpN sites
• Improved quantification of sequence variations at any sequence position
• Easy and improved base calling functionality
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.
PCR Array Data Analysis Tutorial: qPCR Technology Webinar Series Part 3QIAGEN
Using actual PCR Array data, this slidedeck presents an easy-to-use and free web-based data analysis tool to calculate fold-differences in gene expression from your raw real-time PCR threshold cycles. Learn how you can look at your results in different formats, including heat map, scatter, volcano, clustergram and multigroup plot.
This slidedeck presents a simple and accurate real-time PCR system for relevant biological pathway- and disease-focused mRNA and long noncoding RNA (lncRNA) expression profiling. Learn about the stringent performance built into the technology to ensure its sensitivity, specificity, reproducibility and reliability. Application examples are also presented.
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!
New Progress in Pyrosequencing for DNA MethylationQIAGEN
Pyrosequencing is a highly flexible technology that lets you rapidly analyze short- to medium-length sequences fast and quantitatively with high accuracy. The real-time, high-resolution sequence output makes the technology highly suitable for applications including complex mutation analysis, microbial identification and DNA methylation quantification.
The main bottleneck in Pyrosequencing has been limited sequence length, which is critical for some applications. Our new technology, software, and chemistry overcome this bottleneck and give sequence reads that are typically twice as long as those from previous PyroMark systems. The new PyroMark Q24 Advanced system also reduces background noise, improving quantification even at sites distant from the sequencing start. The new system is ideal for applications requiring analysis of longer sequences, such as DNA methylation analysis in epigenetic research, frequency determination in mutation analysis, and various de novo sequencing applications.
In this presentation, we will discuss the following applications and technology improvements:
• DNA methylation analysis at single base resolution at CpG and CpN sites
• Improved quantification of sequence variations at any sequence position
• Easy and improved base calling functionality
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.
Advanced miRNA Expression Analysis: miRNA and its Role in Human Disease Webin...QIAGEN
miRNAs are small functional RNAs, which regulate gene expression post-transcriptionally. The miScript miRNA PCR Array System is a sensitive and reliable technology for detection of mature miRNAs in any laboratory. In this slideshow, the challenges of miRNA data analysis and solutions that the miScript miRNA PCR Arrays provide for researchers interested in identifying miRNA from cells, tissues and FFPE samples are described. You will also learn how to use our GeneGlobe Data Analysis Center to identify miRNAs that may be important in your favorite biological pathway or disease.
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.
RNA Integrity and Quality – Standardize RNA Quality Control QIAGEN
RNA integrity and quality are critical to obtain meaningful and reliable downstream data. This slidedeck details the challenges and considerations of handling RNA samples, the need for quality control analysis and common methods for RNA integrity and quality assessment. The QIAxcel Advanced System will be introduced to automate the process of RNA sample integrity analysis and obtain objective quality measurement. Application data will be presented.
In this slidedeck, the following topics, which are critical steps for efficient and precise gene expression studies using real-time PCR technology, are covered:
• Effect of RNA integrity on real-time PCR results – tips on how to achieve a true RNA profile suitable for real-time PCR studies
• Improved methods for cDNA synthesis, optimized for real-time PCR
• Real-time PCR analysis
• Real-time PCR essentials and background information on different quantification strategies
• SYBR Green real-time PCR – factors influencing specificity
• Introduction to probe technology
• New, fast and efficient real-time PCR solutions
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 of RNA Samples - For Gene-Expression Results you Can Rely onQIAGEN
By their very nature RNA molecules, especially mRNA and regulator RNA, are labile and can be highly unstable and sensitive to heat, UV and RNase contamination. The quality, relevance and scientific impact of gene expression results directly depends on the ability to extract RNA without losing any fraction of interest, while preserving the integrity of the biological information it carries. RNA quality control is thus critical to ensure high-quality results and for turning these results into actionable insights with confidence.
In this webinar, we will introduce you to the main parameters influencing RNA-based assays and their respective impact on downstream applications, discuss how to monitor them and cover the advantages of automation for quality control along complex workflows.
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.
The importance of controls and novel solutions for successful real-time qPCRQIAGEN
The increasing demand for streamlined, monitored and ultrafast qPCR procedures requires high-performance, real-time quantitative RT and PCR chemistries. Particularly, procedures utilizing generic kits for gene expression analysis should include in-process safety measures to avoid variables and control accuracy of procedures and results. This slidedeck presents innovative solutions for one-step and two-step RT-PCR that significantly enhance performance and reliability in qRT-PCR. The new QuantiNova kit family offers a combination of various integrated safety features to remove variables and prevent artifacts. Internal control RNA, removal of genomic DNA, room temperature set-up capability for RT-PCR and a built-in visual pipetting control verify accurate procedures, ensuring reliable gene expression profiling.
This slidedeck explains the principles of the technologies and shows data demonstrating performance in qRT-PCR. Find out how you can verify accurate performance in qRT-PCR and improve your results!
Critical Steps for Real-Time PCR Analysis: Tips and Solutions to Achieve Effi...QIAGEN
In this slidedeck, we cover the following topics which are critical steps for efficient and precise gene expression studies using real-time PCR technology:
1) Effect of RNA integrity on real-time PCR results – tips to achieve a true RNA profiling suitable for real-time PCR studies
2) Improved methods for cDNA synthesis, optimized for real-time PCR
3) Real-time PCR analysis:
• Real-time PCR essentials and background information on different quantification strategies
• SYBR Green real-time PCR – factors influencing specificity
• Introduction to probe technology
• New, fast and efficient real-time PCR solutions
RNA integrity and quality - Standardize RNA quality controlQIAGEN
RNA integrity and quality are critical to obtain meaningful and reliable downstream data. This slidedeck details the important considerations and critical factors in RNA preparation. It also highlights the need for quality control analysis and common methods for RNA integrity and quality assessment.
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.
At the heart of every successful discovery lie the seeds of innovation. At QIAGEN, we are constantly developing new methods that allow researchers to gain forward momentum with their research. Whether you’re studying gene expression or performing viral RNA analysis, the success of your experiment depends on the ability to analyze your sample with the highest standards of sensitivity and specificity so that you can have confidence in your data. To help you generate valuable insights from gene expression profiling and viral RNA analysis, we introduce the brand new QIAGEN OneStep Ahead RT-PCR Kit – the first hot start reverse transcriptase kit on the market. Continuing the success story of its first-generation predecessor (QIAGEN OneStep RT-PCR Kit), the QIAGEN OneStep Ahead RT-PCR Kit is equipped with compelling new features that afford maximum convenience and ease of use, while delivering unmatched sensitivity and specificity. With a total reaction time of 1 hour, higher sequence accuracy and the ability to amplify amplicons of up to 4 kb without tedious optimization, you can get one step closer to publishing your findings with this new solution. For increased convenience, the kit comes in an all-in-one tube format along with a built-in pipetting control. Stay one step ahead of your peers and make significant advances in your research with the QIAGEN OneStep Ahead RT-PCR Kit! In this slidedeck, we introduce the new kit in detail and discuss its features and benefits.
Gene Expression Assay Performance Guaranteed With the TaqMan® Assays QPCR Gua...Thermo Fisher Scientific
Real-time or quantitative PCR (qPCR) is one of the most powerful and sensitive techniques available for gene expression analysis. It is used for a broad range of applications, including quantification of gene expression, measuring RNA interference, biomarker discovery, pathogen detection, and drug target validation. When studying gene expression with qPCR, scientists usually investigate changes—increases or decreases—in the quantity of particular gene products or a set of gene products. Investigations typically evaluate gene response to biological conditions such as disease states, exposure to pathogens or chemical compounds, the organ or tissue location, or cell cycle or differentiation status.
Real-time PCR for the quantification of gene expression using the 5’ nuclease assay with TaqMan® probes has become a standard method in basic and clinical research.
http://owl.li/dgR59
RNase H2-dependent PCR (rhPCR) is a powerful method for increasing PCR specificity and eliminating primer-dimers by using blocked primers and a thermostable RNase H2 from Pyrococcus abyssi (P. abyssi). Primers will only support extension and replication after the blocked portion is cleaved. Cleavage by the RNase H2 enzyme occurs only when primers are bound to their complementary target sequence, thus providing increased specificity. Also, the thermostability of P. abyssi RNase H2 provides a “hot start” capability to the reaction. In this presentation, Dr Joseph Dobosy (senior research scientist in the molecular genetics research division of IDT) gives a detailed explanation of the rhPCR mechanism, offer tips on how to design assays using this powerful technology, and discuss examples of applications that benefit from rhPCR.
Profile Multiple Cytokines and Chemokines Simultaneously with Very High Sensi...QIAGEN
Learn how to profile multiple cytokines and chemokines simultaneously with very high sensitivity and specificity using the standard ELISA reader. Available in different formats to suit your research needs such as single-analyte, multi-analyte or custom mix-n-match format for human, mouse and rat.
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.
Advanced miRNA Expression Analysis: miRNA and its Role in Human Disease Webin...QIAGEN
miRNAs are small functional RNAs, which regulate gene expression post-transcriptionally. The miScript miRNA PCR Array System is a sensitive and reliable technology for detection of mature miRNAs in any laboratory. In this slideshow, the challenges of miRNA data analysis and solutions that the miScript miRNA PCR Arrays provide for researchers interested in identifying miRNA from cells, tissues and FFPE samples are described. You will also learn how to use our GeneGlobe Data Analysis Center to identify miRNAs that may be important in your favorite biological pathway or disease.
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.
RNA Integrity and Quality – Standardize RNA Quality Control QIAGEN
RNA integrity and quality are critical to obtain meaningful and reliable downstream data. This slidedeck details the challenges and considerations of handling RNA samples, the need for quality control analysis and common methods for RNA integrity and quality assessment. The QIAxcel Advanced System will be introduced to automate the process of RNA sample integrity analysis and obtain objective quality measurement. Application data will be presented.
In this slidedeck, the following topics, which are critical steps for efficient and precise gene expression studies using real-time PCR technology, are covered:
• Effect of RNA integrity on real-time PCR results – tips on how to achieve a true RNA profile suitable for real-time PCR studies
• Improved methods for cDNA synthesis, optimized for real-time PCR
• Real-time PCR analysis
• Real-time PCR essentials and background information on different quantification strategies
• SYBR Green real-time PCR – factors influencing specificity
• Introduction to probe technology
• New, fast and efficient real-time PCR solutions
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 of RNA Samples - For Gene-Expression Results you Can Rely onQIAGEN
By their very nature RNA molecules, especially mRNA and regulator RNA, are labile and can be highly unstable and sensitive to heat, UV and RNase contamination. The quality, relevance and scientific impact of gene expression results directly depends on the ability to extract RNA without losing any fraction of interest, while preserving the integrity of the biological information it carries. RNA quality control is thus critical to ensure high-quality results and for turning these results into actionable insights with confidence.
In this webinar, we will introduce you to the main parameters influencing RNA-based assays and their respective impact on downstream applications, discuss how to monitor them and cover the advantages of automation for quality control along complex workflows.
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.
The importance of controls and novel solutions for successful real-time qPCRQIAGEN
The increasing demand for streamlined, monitored and ultrafast qPCR procedures requires high-performance, real-time quantitative RT and PCR chemistries. Particularly, procedures utilizing generic kits for gene expression analysis should include in-process safety measures to avoid variables and control accuracy of procedures and results. This slidedeck presents innovative solutions for one-step and two-step RT-PCR that significantly enhance performance and reliability in qRT-PCR. The new QuantiNova kit family offers a combination of various integrated safety features to remove variables and prevent artifacts. Internal control RNA, removal of genomic DNA, room temperature set-up capability for RT-PCR and a built-in visual pipetting control verify accurate procedures, ensuring reliable gene expression profiling.
This slidedeck explains the principles of the technologies and shows data demonstrating performance in qRT-PCR. Find out how you can verify accurate performance in qRT-PCR and improve your results!
Critical Steps for Real-Time PCR Analysis: Tips and Solutions to Achieve Effi...QIAGEN
In this slidedeck, we cover the following topics which are critical steps for efficient and precise gene expression studies using real-time PCR technology:
1) Effect of RNA integrity on real-time PCR results – tips to achieve a true RNA profiling suitable for real-time PCR studies
2) Improved methods for cDNA synthesis, optimized for real-time PCR
3) Real-time PCR analysis:
• Real-time PCR essentials and background information on different quantification strategies
• SYBR Green real-time PCR – factors influencing specificity
• Introduction to probe technology
• New, fast and efficient real-time PCR solutions
RNA integrity and quality - Standardize RNA quality controlQIAGEN
RNA integrity and quality are critical to obtain meaningful and reliable downstream data. This slidedeck details the important considerations and critical factors in RNA preparation. It also highlights the need for quality control analysis and common methods for RNA integrity and quality assessment.
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.
At the heart of every successful discovery lie the seeds of innovation. At QIAGEN, we are constantly developing new methods that allow researchers to gain forward momentum with their research. Whether you’re studying gene expression or performing viral RNA analysis, the success of your experiment depends on the ability to analyze your sample with the highest standards of sensitivity and specificity so that you can have confidence in your data. To help you generate valuable insights from gene expression profiling and viral RNA analysis, we introduce the brand new QIAGEN OneStep Ahead RT-PCR Kit – the first hot start reverse transcriptase kit on the market. Continuing the success story of its first-generation predecessor (QIAGEN OneStep RT-PCR Kit), the QIAGEN OneStep Ahead RT-PCR Kit is equipped with compelling new features that afford maximum convenience and ease of use, while delivering unmatched sensitivity and specificity. With a total reaction time of 1 hour, higher sequence accuracy and the ability to amplify amplicons of up to 4 kb without tedious optimization, you can get one step closer to publishing your findings with this new solution. For increased convenience, the kit comes in an all-in-one tube format along with a built-in pipetting control. Stay one step ahead of your peers and make significant advances in your research with the QIAGEN OneStep Ahead RT-PCR Kit! In this slidedeck, we introduce the new kit in detail and discuss its features and benefits.
Gene Expression Assay Performance Guaranteed With the TaqMan® Assays QPCR Gua...Thermo Fisher Scientific
Real-time or quantitative PCR (qPCR) is one of the most powerful and sensitive techniques available for gene expression analysis. It is used for a broad range of applications, including quantification of gene expression, measuring RNA interference, biomarker discovery, pathogen detection, and drug target validation. When studying gene expression with qPCR, scientists usually investigate changes—increases or decreases—in the quantity of particular gene products or a set of gene products. Investigations typically evaluate gene response to biological conditions such as disease states, exposure to pathogens or chemical compounds, the organ or tissue location, or cell cycle or differentiation status.
Real-time PCR for the quantification of gene expression using the 5’ nuclease assay with TaqMan® probes has become a standard method in basic and clinical research.
http://owl.li/dgR59
RNase H2-dependent PCR (rhPCR) is a powerful method for increasing PCR specificity and eliminating primer-dimers by using blocked primers and a thermostable RNase H2 from Pyrococcus abyssi (P. abyssi). Primers will only support extension and replication after the blocked portion is cleaved. Cleavage by the RNase H2 enzyme occurs only when primers are bound to their complementary target sequence, thus providing increased specificity. Also, the thermostability of P. abyssi RNase H2 provides a “hot start” capability to the reaction. In this presentation, Dr Joseph Dobosy (senior research scientist in the molecular genetics research division of IDT) gives a detailed explanation of the rhPCR mechanism, offer tips on how to design assays using this powerful technology, and discuss examples of applications that benefit from rhPCR.
Profile Multiple Cytokines and Chemokines Simultaneously with Very High Sensi...QIAGEN
Learn how to profile multiple cytokines and chemokines simultaneously with very high sensitivity and specificity using the standard ELISA reader. Available in different formats to suit your research needs such as single-analyte, multi-analyte or custom mix-n-match format for human, mouse and rat.
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.
Technical Guide to Qiagen PCR Arrays - Download the GuideQIAGEN
Total RNA discovery with RT2 and miScript PCR Arrays : Explore the RNA universe - Whatever your destination within the RNA universe, QIAGEN will help you get there. The miRNeasy kits deliver pure, high-quality total RNA from a broad range of samples. The RT2 and miScript PCR arrays are a complete solution both for focused analysis of gene and microRNA expression and for validation of microarray and RNA sequencing experiments. Together with the powerful analytics tools of GeneGlobe® and QIAGEN Ingenuity® Pathway Analysis, these products give you a smooth path from your sample to high-quality results.
Genomics research and discovery has led to a large increase of reported single nucleotide polymorphisms (SNPs). From 2006 to 2017, the number of refSNPs in the NCBI dbSNP database has increased 13-fold. Many polymorphisms can be linked to disease susceptibility and responses to chemical therapies. Other polymorphisms are used as trait identifiers in livestock and plants. Being able to inexpensively and accurately determine the genotype in high-throughput fashion, with low sample input is a critical need in current, large-scale screening efforts. In this presentation, we present a novel, probe-based, PCR genotyping solution that possesses the universal cycling conditions, strong signal generation, and benchtop reaction stability needed for high-throughput screening. We also present the mechanism and unique technical advantages of using the rhAmp SNP Genotyping System, and we will illustrate how easy it is to generate high quality genotyping data.
Single-nucleotide polymorphisms (SNPs) provide important information about the biology and evolution of different organisms. SNPs may also help predict an individual’s response to certain drugs, susceptibility to environmental factors, and risk of developing particular diseases providing valuable insight into pathophysiology of the human condition. As a result, SNPs with important functional roles often become subjects for high-throughput experiments.
In this webinar, Daniel Tsang provides an overview of genotyping using real-time PCR (qPCR) technology, including challenges and ways to overcome these challenges. He presents a novel qPCR-based genotyping solution, the rhAmp™ SNP Genotyping System, along with its advantages in genotyping, details on cluster separation, as well as solutions to improve the calling accuracy and confidence of making genotype calls.
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.
MOLECULAR TOOLS IN DIAGNOSIS AND CHARACTERIZATION OF INFECTIOUS DISEASES tawheedshafi
The future of the molecular diagnostics of infectious diseases will undoubtedly be focused on a marked increase in the amount of information detected with remarkably simplified, rapid platforms that will need complex software analysis to resolve the data for use in clinical decision-making.
Digital RNAseq for Gene Expression Profiling: Digital RNAseq Webinar Part 2QIAGEN
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. In this webinar we will cover, in depth, the 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.
3. Central Dogma of Molecular Biology
Transcription
DNA
Genome
10,000 20,000 Genes
Translation
mRNA
Transcriptome
Protein
Proteome
~ 50,000 proteins
Sample & Assay Technologies
4. Gene Expression (mRNA) Regulates Biology
Biological Processes, such as Apoptosis, Cell
Cycle, Inflammation, Signal Transduction, etc.,
can be identified by specific patterns of gene
expression (pathway-focused).
Sample & Assay Technologies
5. Differential Gene Expression Analysis
Q: How to Assess the Expression of
Different mRNAs in a Sample and
Compare it across multiple samples?
A:
Sample & Assay Technologies
7. Table of Contents
1. Introduction to PCR Arrays
2. How PCR Arrays Work
3. Performance Data from PCR Arrays
4. Research Applications for PCR Arrays
Sample & Assay Technologies
8. PCR Arrays for ALL Biomedical Researchers
Cancer and Apoptosis
Cytokines & Inflammation
Development & Stem Cells
Apoptosis
Inflammatory Cytokines
Stem Cells
Cell Cycle
Th17 for Inflammation
WNT Signaling / Notch Signaling
Human miRNA Array (NEW!)
Common Cytokines / Chemokines
Terminal Differentiation Markers
Breast Cancer & Estrogen Receptor
Inflammasomes - NEW!
TGFβ / BMP Signaling
Tumor Metastasis
NF-kB Signaling Pathway
Endothelial Cell Biology
Epithelial-to-Mesenchymal Transition
Th1-Th2-Th3
Osteogenesis
Angiogenesis
TNF Ligands
Growth Factors
Cancer Drug Resistance
Toll-like Receptors
ECM & Adhesion
Signal Transduction
Toxicology & Drug Metabolism
Neuroscience
Signal Transduction PathwayFinder
Drug Metabolism
Neuroscience Ion Channels
NFkB Signaling
Drug Phase I Enzymes
Neurotransmitter Receptors
Jak / Stat Signaling
Drug Transporters
Neurotrophins & Receptors
DNA Damage Signaling
Oxidative Stress
Neurogenesis and Neural Stem Cell
Insulin Signaling
Stress & Toxicity
MAP Kinase Signaling
Other Diseases
Custom PCR Arrays (H/M/R/Q/D/F)
cAMP / Calcium Signaling
Atherosclerosis
96-Well, 384-Well Plate
p53 Signaling
Diabetes
100-Well Disc, 96x96 Chip
** Over 100 Pathway-Powered PCR Arrays Available**
Sample & Assay Technologies
9. Primer Assays – Laboratory Verification Data
Melting/Dissociation Curve
• Single Peak
Amplification Plot
• PCR Efficiency
Sample & Assay Technologies
10. How Genes on PCR Arrays Are Selected
• Biologically relevant gene content
• Not simply biochemical pathways or kinase
cascades
• Published association with the biological or
disease pathway gathered from overlapping
sources, including:
• Multiple Publicly Accessible Databases
• Text Mining Relevant Literature
• Technically relevant gene content
• Use genes that are regulated at the mRNA
level
• Specific feedback from thought leaders
Genes in Signaling Pathways
Genes with High Relevance
Sample & Assay Technologies
12. Complete PCR Array System – Bench Verified
• The entire system has been optimized and bench-verified and
bench
we guarantee their performance when used together!
• RNA isolation kits (verified)
• RT2 Profiler PCR Arrays - Bench verified
• RT2 1st Strand cDNA synthesis kit
–
–
–
–
Efficient genomic DNA removal step
Built-in reverse transcription control to monitor sample quality
A hybrid method of random primers and Oligo dT
Fast (20 minutes); High-throughput available
• RT2 SYBR Green Master Mixes
– Unique buffer system to ensure PCR specificity and efficiency
• Free PCR Array Data Analysis software
– Web & Excel based
Sample & Assay Technologies
13. Table of Contents
1. Introduction to PCR Arrays
2. How PCR Arrays Work
3. Performance Data from PCR Arrays
4. Research Applications for PCR Arrays
Sample & Assay Technologies
20. PCR Array Compatible RT-PCR Instruments
96-, 100-, 384-, and 96x96 Well Formats
•
•
•
96-Well Blocks: 7000, 7300, 7500, 7700,
ViiA 7
FAST 96-Well Blocks: 7500, 7900HT, Step
One Plus, ViiA 7
FAST 384-Well Block: 7900HT, ViiA 7
•
•
iCycler, MyiQ, MyiQ2, iQ5, CFX96, CFX384
Opticon, Opticon 2, Chromo 4
•
Mastercycler ep realplex 2/2S/4/4S
•
BioMark
•
Rotor-Gene Q, Rotor-Gene 6000
•
LightCycler 480
•
Mx3000p, Mx3005p, Mx4000p
•
TP-800
New Instrument
Set-up protocol available
• No real time PCR machine? PCRArray Service from SABio
Sample & Assay Technologies
21. RT2 Profiler™ PCR Array Data Analysis
• FREE Complete & Easy Data Analysis with Web-Based Software
• From Raw Ct Values to Fold Change Results in Multiple Analysis Formats
Volcano Plot
Scatter Plot
Clustergram
3-D Histogram
Register for PCR Array Data Analysis webinar to learn more!
Sample & Assay Technologies
22. Table of Contents
1.
Introduction to the PCR Arrays
2.
How PCR Arrays Work
3.
Performance Data from PCR Arrays
–
All individual PCR Primer Assays are wet-bench validated for:
• Sensitivity
– Enhanced Sensitivity with PreAMP Technology
• Specificity
• Amplification Efficiency
• Consistency
4.
Research Applications for PCR Arrays
Sample & Assay Technologies
23. Superb Sensitivity
See More Genes with Less RNA
• Human Inflammatory Cytokines &
Receptors Array used to analyze uninduced cells. Genes represented at
very low copy numbers.
• 25 ng - 5 µg total RNA per 96-well plate
• 99% of genes detectable with > 500 ng
• > 80% of genes detectable with 25 ng
• Very sensitive assay!
Human Inflammatory Cytokines & Receptors
PCR Array tested with Universal Reference RNA
Conclusion: See gene expression as rare as UN-induced cytokines.
Sample & Assay Technologies
24. Exquisite Specificity
• Assays on Human TGFβ &
BMP Signaling Pathway
PCR Array used to analyze
BMP genes in Universal
Reference RNA.
• Single peak dissociation
curves
• Single gel bands of
predicted size
• High specificity for genes
difficult to design specific
primers for.
Conclusion: Each well in a PCR Array detects a single genespecific product.
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25. High Reproducibility
Reproducibility Among Different Users
Reproducibility Among Different Instruments
Human Drug Metabolism PCR Array in Universal Reference RNA
Conclusion: Same raw threshold cycle data can be obtained for
the same samples even from different end users at different
times or using different instruments.
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26. Uniform Amplification Efficiency
Representative set of 500 out of > 4,000 assays used in PCR Arrays
Conclusion: Accurate and reliable ∆∆Ct results
are guaranteed.
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27. Wide Dynamic Range
Human Acetylcholine Receptor α5
Conclusion: Wide dynamic range to
accommodate varying levels of initial
gene expression.
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28. Replicates: Technical & Biological
Technical
• Not necessary
Save Time & Money
• Reproducibility of the PCR Arrays is very high
• Results demonstrate that what you are seeing is a result
of biology, not technique.
• RTC & PPC show technical reproducibility on each plate,
and comparable across plates.
Biological
• Needed to verify the results are a result of biology
• At least 3 replicates per sample for statistical analysis
• p values
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29. Table of Contents
1. Introduction to the PCR Arrays
2. How PCR Arrays Work
3. Performance Data from PCR Arrays
RT2 PreAMP
Pathway-Powered PreAMPlification
4. Research Applications for PCR Arrays
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30. Small or Fixed Samples:
A Challenge for RT-PCR
• Samples containing less than 25 ng
• Sources include:
• Stem Cells
• Fine Needle Biopsies/ Aspirations
• Laser Capture Microdissection
• Sorted Samples (FACS, selection markers, GFP)
• Formalin-fixed Paraffin-embedded samples
• Sources include:
• Research animals
• Clinical samples
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31. PCR Arrays with PreAMP – Increased Sensitivity
See More Genes with Even Less RNA – Pathway-Focused
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32. How RT2 PCR Arrays Work
Cells & Tissues RNA (25ng- 5ug)
gDNA Elimination Step
~45 min
First Strand Synthesis
1st strand cDNA
+ SYBR Green Master Mix
2 hours
Real-time PCR Detection of 89
Gene-specific Amplification on
RT2 Profiler PCR Array
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33. How RT2 PreAMP Technology Works
Tandem PCR-based Technology
Cells & Tissues RNA (1ng-100ng); FFPE RNA (100ng)
gDNA Elimination Step
~45 min
First Strand Synthesis
(1/4 of RT product i.e. 5µL)
µ
1st strand cDNA
~40 min
+ PreAMP Primer Mix
and PreAMP Master Mix
PreAMP PCR Step
(Nano:12 Cycles
FFPE: 8 Cycles)
Amplified cDNA
~20 min
Total time:< 2 hr
2 hours
+ Side Reaction Reducer
(Exonuclease I)
Eliminate Residual
PreAMP Primers
Clean Amplified cDNA
+ SYBR Green Master Mix
Real-time PCR Detection of 89
Gene-specific Amplification on
RT2 Profiler PCR Array
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34. Essentials of RT2 PreAMP Technology
96-well Pathway-Powered PCR Array
• 84 Genes of Interest
• 5 Housekeeping Genes
• 7 Controls
Each PreAMP Primer Mix
• 84 Genes of Interest
• 1 primer set / gene
• 84 primer sets combined
RT2 PreAMP: Pathway-Focused Gene Amplification
• Faithfully Amplify ONLY those Genes Appearing
on Your Array
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35. RT2 PreAMP: Increased Positive Call Rate
• Sample: Different amounts of Human universal total RNA were reverse transcribed to cDNA with (red bars) or without
PreAMPlification (blue bars) using RT² PreAMP PCR Master Mix and RT² PreAMP cDNA Synthesis Primer Mix.
• PCR Array: Human Inflammatory Cytokines and Receptors PCR Array
• Unamplified: As little as 25 ng total RNA before PreAMP yields greater than an 80% positive call
• Nano PreAMPlified: Increased sensitivity observed with RT2 Profiler PCR Array, allowing analysis of total RNA as little as
1 ng with a positive call rate greater than 90%.
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36. ∆Ct-PreAMP cDNA
RT2 PreAMP: Unbiased Amplification Process
R2=0.95; Slope=1.01
∆Ct-Unamplified cDNA
• Sample: Human Liver Tumor RNA was reverse transcribed to cDNA with or without PreAMPlification using RT² PreAMP PCR
Master Mix and RT² PreAMP cDNA Synthesis Primer Mix.
• PCR Array: Human Cancer PathwayFinder
• Unamplified cDNA: 500 ng total RNA
• Nano PreAMPlified cDNA: 5 ng total RNA
• Unamplified cDNA and Nano PreAMPlified cDNA, when reverse transcribed and analyzed on a RT2 Profiler PCR Array, yield
similar Ct values for each gene under observation.
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37. Table of Contents
1. Introduction to the PCR Arrays
2. How PCR Arrays Work
3. Performance Data from PCR Arrays
4. Research Applications for PCR Arrays
Sample & Assay Technologies
38. Publish in High Impact Factor Journals
SABiosciences.com/support_publication.php#pcrarray
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39. Application Example 1: Cytokine Profiling
Common Cytokine PCR Array: Identified 23 Up-Regulated and 6
Down-Regulated Genes Following PMA and Ionomycin Treatment.
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40. Application Example 2: Tumor Analysis
ECM & Cell Adhesion PCR Arrays: Revealed Up- and
Down- Regulated Genes in Breast Cancer.
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41. Application Example 3: Toxicity Studies
Stress & Toxicity PathwayFinder™ PCR Array: Uncovered Distinct
Gene Expression Profiles Associated with Liver Toxicity Caused by 3 PPARγ
γ
Agonists (Actos, Avandia, and Rezulin).
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42. Create Your Own PCR Arrays:
Custom Formats
• Choose any gene from the Human, Mouse, Rat, Drosophila, Rhesus macaque, or Dog Genomes
96-well
384-well
•
Analyze the Genes Most
Important To Your Research
•
Available for any real-time PCR
instrument.
•
Wet-Bench Validated design &
performance.
•
2 to 3 week turn around time
first order
•
2 to 3 DAY turn around on reorder
•
12 Genes : 8 Samples
Same Data Analysis as
Catalogued Arrays
24 Genes : 4 Samples
32 Genes : 3 Samples
48 Genes : 2 Samples
96 Genes : 1 Sample
Custom 100-discs also available
# of
samples x 4
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43. What’s Next - After PCR Arrays
•
Validate results with more sample and focused set of genes
– RT2 Single Primer Assays
– RT2 Custom PCR Arrays
•
Identify Transcription Factors Regulating your Gene
– SureFind™ Transcriptome PCR Arrays
•
MicroRNA studies
– RT2 MicroRNA PCR Arrays and assays
•
Assess Biological Impact
– Cignal™ Reporters
•
Analyze interaction between DNA & nuclear proteins
– ChampionChIP Chromatin Immunoprecipitation PCR Arrays
•
Knockdown Analysis
– SureSilencing shRNA Plasmids
•
Quantify secreted proteins in blood plasma sera
– ELISArrays
Sample & Assay Technologies
44. RT2 PCR Array System SUMMARY
•
•
•
•
Complete System
• RNA isolation kits, PCR Arrays, master mix, first strand kit, and free data analysis
software
• Systematic controls to monitor sample quality, RT and PCR efficiency
• 3 hours from RNA to data analysis
Breadth of Pathway Content
• Profile the genes and pathways that you really care about
• More than 100 pathways for human, mouse, and rat
• Customizable by gene, set, or pathway
Superb Real-Time PCR Performance
• Reproducibility, Sensitivity, Specificity, Reliability
Broad Application
• Cancer, Immunology, Neuroscience, Stem Cells & much more
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46. PCR Arrays
EmPower Your Pathway Analysis
Experience PCR Array Performance
Try them today!
Questions?
Contact Technical Support 9 AM – 6 PM Eastern M – F
Contact: 1-888-503-3187 OR support@SABiosciences.com
Shankar Sellappan, Ph.D. (Global Product Manager):
Shankar.Sellappan@QIAGEN.com
Thousands of scientists have
discovered the power of PCR Arrays.
Join them on the road to success!
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