Successful Validation of RNA Targets: Improve your gene expression analysis with our target validation workflow
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RNA target validation workflow using high-performance technologies for reliable and reproducible results.
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Enabling RNA-Seq With Limited RNA Using Whole Transcriptome Amplification
RNA-Seq was developed to perform transcriptome profiling and provides a highly precise measurement of expression levels of transcripts and their isoforms. Normally, RNA-Seq analysis requires at least 500 ng –1 μg of total RNA. When working with small biopsies, single cells (such as circulating tumor cells), or other limited material, whole transcriptome amplification (WTA) is normally required. Various WTA methods overcome limited RNA availability and enable transcriptome analysis from limited material or even single cells. In standard PCR-based WTA procedures, however, bias from uneven coverage of cDNA regions with high GC or AT content or amplification errors can lead to the loss of transcripts and wrong variant calling. Here, we compare a standard RNA-Seq library preparation method and the REPLI-g RNA library protocol. The REPLI-g procedure is a PCR-free protocol to efficiently generate RNA-Seq libraries from small amounts of RNA or a single cell in 6.5–7 hours. The REPLI-g protocol uses whole transcriptome amplification based on multiple displacement amplification (MDA), combined with an efficient library adaptor ligation procedure, to prepare RNA-Seq libraries from small RNA amounts. The procedure demonstrates high fidelity, minimal bias and retention of sample‘s transcriptional profile. Compared to standard RNA-Seq library prep, the REPLI-g protocol demonstrates similar reproducibility and sensitivity in transcript detection.
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• A ligation-based library construction method using a GeneRead Library Prep Kit (QIAGEN)
• A ‘tagmentation’-based method using Nextera DNA Sample Prep Kit (Illumina), which simultaneously fragments and tags DNA.
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In this webinar, we will introduce you to the main sample quality parameters and their respective impact on downstream applications, discuss how to monitor them and cover the advantages of automating quality control along complex workflows.
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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.
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RNA profiling is a powerful technique for understanding cellular origins and disease states. Recent studies in a variety of diseases have revealed RNA signatures that are excellent biomarker candidates for understanding disease status and predicting progression.
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Next generation sequencing & microarray-- Genotypic Technology
Greetings from Genotypic Technology, Bangalore (www.genotypic.co.in). We are a 13 year old genomics and bioinformatics company ( 65+ employees, Service. Products and R & D) based in Bangalore, India, primarily working on applications of Microarrays and Next Generation Sequencing in Human Health and Disease, including in Molecular Diagnostics, Prognosis, Therapeutics, Vaccine Research, Basic Science Research, Veterinary Science, Agriculture, Industrial Biotechnology, Microbial Genetics and more.
Our major strength is in customized genomics solutions, particularly in your field, we can develop panel of markers for specific diseases, optimize, validate and help commercialize on open platforms or specific instrument platforms- in microarrays and sequencing based methods/ assays. We can also use genomic markers to aid in treatment of specific disease using personalized medicine approaches. All this can be done on a comprehensive end-to-end manner in our company as we have a very good blend of molecular biology and bioinformatics with totally 6 Ph.Ds. We work closely with Agilent's R &D as their partner.
Technical Guide to Qiagen PCR Arrays - Download the Guide
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Introduction to real-Time Quantitative PCR (qPCR) - Download the slides
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 field-deployable RT-PCR system performs equivalently to real-time RT-PCR in...
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High data quality and accuracy are recognized characteristics of Sanger re-sequencing projects and are primary reasons that next generation sequencing projects compliment their results by capillary electrophoresis data validation. We have developed an on-line tool called Primer Designer™ to streamline the NGS-to-Sanger sequencing workflow by taking the laborious task of PCR primer design out of the hands of the researcher by providing pre-designed assays for the human exome. The primer design tool has been created to enable scientists using next generation sequencing to quickly confirm variants discovered in their work by providing the means to quickly search, order and receive suitable pre-designed PCR primers for Sanger sequencing. Using the Primer Designer™ tool to design M13-tailed and non-tailed PCR primers for Sanger sequencing we will demonstrate validation of 28-variants across 24-amplicons and 19-genes using the BDD, BDTv1.1 and BDTv3.1 sequencing chemistries on the 3500xl Genetic Analyzer capillary electrophoresis platform.
Analyzing Fusion Genes Using Next-Generation Sequencing
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In this webinar, Dr. Raed Samara will cover:
1. Fusion genes: what they are and a historical perspective
2. Fusion gene detection: the current status
3. RNA sequencing vs. digital RNA sequencing
4. How to detect and accurately quantify novel fusion genes in your sample
Identity testing by NGS as a means of risk mitigation for viral gene therapies
Watch the presentation of this webinar here: https://bit.ly/3RijkHC
Detailed description:
Imagine you’ve just completed a manufacturing run for your viral vector. Identity testing is performed to confirm the vector sequence. But when the results come back the data reveals unexpected sequence variants! With an appropriate risk mitigation testing strategy, this situation can be prevented.
The situation described above is not hypothetical, and happens more that you think, costing valuable time and resources.
Investigatory testing has shown that sequence variants present in starting materials (e.g. plasmids) are likely to make their way to the final product. Adequate identification of low-level variants with an appropriately sensitive method is critical in ensuring the quality of the final product. A risk-based testing strategy, in the context of identity, for viral vector manufacturing will be presented, focusing on key testing points. NGS assays for identity and variant detection will be highlighted due to their extremely sensitive nature compared to traditional approaches.
In this webinar, we'll explore:
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• NGS applications for identity testing as compared to traditional methods
• A case study on the impact of not establishing a proper risk-based testing strategy
Presented by: Bradley Hasson, Director of Lab Operations for NGS Services
Identity testing by NGS as a means of risk mitigation for viral gene therapies
Watch the presentation of this webinar here: https://bit.ly/3RijkHC
Detailed description:
Imagine you’ve just completed a manufacturing run for your viral vector. Identity testing is performed to confirm the vector sequence. But when the results come back the data reveals unexpected sequence variants! With an appropriate risk mitigation testing strategy, this situation can be prevented.
The situation described above is not hypothetical, and happens more that you think, costing valuable time and resources.
Investigatory testing has shown that sequence variants present in starting materials (e.g. plasmids) are likely to make their way to the final product. Adequate identification of low-level variants with an appropriately sensitive method is critical in ensuring the quality of the final product. A risk-based testing strategy, in the context of identity, for viral vector manufacturing will be presented, focusing on key testing points. NGS assays for identity and variant detection will be highlighted due to their extremely sensitive nature compared to traditional approaches.
In this webinar, we'll explore:
• Regulatory requirements for identity testing
• NGS applications for identity testing as compared to traditional methods
• A case study on the impact of not establishing a proper risk-based testing strategy
Presented by: Bradley Hasson, Director of Lab Operations for NGS Services
Arraygen brochure
ArrayGen is a global genomics service provider company which is a one stop solution for Genomics Bioinformatics algorithm development and data analysis. ArrayGen's focus on Genomics, NGS data analysis, custom array design, microarray data analysis thus cover a vast majority of Bioinformatics applications in the life sciences sector.
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Presentation from the Life Technologies booth at PAG
Presentation from the Life Technologies booth at PAG
Accelerate Your Discovery with QIAGEN Service Solutions for Biomarker Researc...
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PCR Primer Design Tool for Sanger Sequencing Confirmation
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Identity testing by NGS as a means of risk mitigation for viral gene therapies
Identity testing by NGS as a means of risk mitigation for viral gene therapies
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- 1. Accurate gene expression results can be difficult to achieve because technical artifacts can mimic biological variability. Successful validation of promising RNA targets using qPCR requires robust, sensitive and standardized tools to verify technical accuracy and determine biological relevance. Precise and reliable qPCR places high demands on instruments, chemistry and software. QIAGEN experts have developed a new RNA target validation workflow using high-performance technologies for reliable and reproducible results. Remove the risk of introducing technical artifacts that prevent true biological insights: discover a better way to validate your targets. Learn more at www.qiagen.com/gene-expression. Successful Validation of RNA Targets! Improve your gene expression analysis results with our target validation workflow Sample to Insight
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