The document describes an RNA-seq analysis workflow called RNACocktail. It evaluates tools for various RNA-seq analysis tasks like alignment, assembly, quantification, and more. The study finds that performance can vary significantly depending on the specific tool and data set. It then proposes a comprehensive RNACocktail protocol and computational pipeline that achieves high accuracy across different sample types and analysis goals. Validation on multiple samples shows this broad analysis approach can help researchers extract more biologically relevant insights from transcriptomic data.
RNA-Seq Analysis: Everything You Always Wanted to Know...and then somebasepairtech
Computational biologist and Basepair founder, Dr. Amit Sinha (@ausinha) helps viewers navigate the world of RNA-Seq analysis. Topics include: Introduction to RNA-Seq, tools and workflows for analysis, visualization and figures, Q & A. More info at: https://www.basepairtech.com/
Enabling RNA-Seq With Limited RNA Using Whole Transcriptome AmplificationQIAGEN
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.
A series of stringent criteria are applied to implement quality control (QC) of mRNA products in order to guarantee reliability.
https://mrna.creative-biolabs.com/in-silico-mrna-structure-prediction.htm
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.
RNA-Seq Analysis: Everything You Always Wanted to Know...and then somebasepairtech
Computational biologist and Basepair founder, Dr. Amit Sinha (@ausinha) helps viewers navigate the world of RNA-Seq analysis. Topics include: Introduction to RNA-Seq, tools and workflows for analysis, visualization and figures, Q & A. More info at: https://www.basepairtech.com/
Enabling RNA-Seq With Limited RNA Using Whole Transcriptome AmplificationQIAGEN
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.
A series of stringent criteria are applied to implement quality control (QC) of mRNA products in order to guarantee reliability.
https://mrna.creative-biolabs.com/in-silico-mrna-structure-prediction.htm
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.
The application of IVT mRNA is a versatile and promising tool for the delivery of transgenes into desired cells. https://mrna.creative-biolabs.com/custom-mrna-modification.htm
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
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.
5 Tips for Successful qRT-PCR Results InfographicQIAGEN
Market research shows that 66.6% of researchers use qRT-PCR for gene-expression profiling. Clearly, this is a very effective and popular technique for detecting RNA expression levels, but it’s still prone to pitfalls that can sometimes lead to disappointing results.
To help improve your experiments, we’ve made a new infographic with some interesting facts about qRT-PCR and 5 tips to help with your gene expression studies. This will cover everything from experimental design to data analysis.
Improving RNA yield and quality is easy with the right protocols. This trouble shooting guide presents methods to overcome common difficulties in RNA purification. Prevent RNA degradation, improve RNA yields, reduce genomic DNA contamination, and more.
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.
Multicopy reference assay (MRef) — a superior normalizer of sample input in D...QIAGEN
Copy number variations (CNVs) and alterations (CNAs) are a source of genetic diversity in humans and are often pathogenic. Numerous CNVs and CNAs are being identified with various genome analysis platforms, including array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) genotyping platforms, and next-generation sequencing. Independent verification of copy number changes is a critical step. Quantitative real-time PCR (qPCR) is a classic method to verify microarray copy number findings. Traditional copy number assays that use qPCR typically rely on a putative single-copy gene reference assay (e.g., RNase P or TERT) to normalize the DNA input for downstream ΔΔCT-based copy number calculation for comparison to a reference genome. When applied to cancer samples, these single-copy reference assays may no longer be a reliable indicator of DNA input due to the presence of complex chromosome composition (both in chromosome number and structure). To meet the need for an accurate DNA input normalizer, especially for heterogeneous tumor samples, QIAGEN developed a multicopy reference (MRef) assay for real-time PCR copy number analysis. This assay, in conjunction with QIAGEN’s greater than 10 million genomewide copy number assays and pathway- and disease-focused copy number PCR arrays (Figure 1), provides a successful solution for copy number analysis. This article will address the assay design considerations, development, and performance of this multicopy reference (MRef) assay.
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!
RT2 Profiler PCR Arrays: Pathway-focused Gene Expression Profiling with qRT-P...QIAGEN
This paper evaluates the performance of the newest technique for monitoring the expression of a panel of pathway- or disease-specific genes: the RT2 Profiler PCR Array System. The RT2 Profiler PCR Array System combines the quantitative performance of SYBR® Green real-time PCR with the multiple-gene profiling capabilities of a microarray.
The RT2 Profiler PCR Array is a 96- or 384-well plate containing RT2 qPCR Primer Assays for a set of 84 related genes, plus 5 housekeeping genes and 3 controls. The complete system includes an instrument-specific master mix and an optimized first strand synthesis kit. This paper presents experimental data showing that RT2 Profiler PCR Arrays have the sensitivity, reproducibility, and specificity expected from real-time PCR techniques. As a result, this technology brings focused gene expression profiling to any biological laboratory setting with a real-time PCR instrument.
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 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
PCR - From Setup to Cleanup: A Beginner`s Guide with Useful Tips and Tricks -...QIAGEN
This End-Point PCR Beginner´s Guide will not only give you a comprehensive overview of tools and techniques to help you to get the most out of your samples, but also give you information on dedicated solutions and complete workflows on multiplex PCR and PCR fragment analysis.
Course: Bioinformatics for Biomedical Research (2014).
Session: 2.3- Introduction to NGS Variant Calling Analysis.
Statistics and Bioinformatisc Unit (UEB) & High Technology Unit (UAT) from Vall d'Hebron Research Institute (www.vhir.org), Barcelona.
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.
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!
CD Genomics provides a fast, one-stop bacterial RNA sequencing solution from the quality control of sample to comprehensive data analysis. Please contact us for more information and a detailed quote.
The application of IVT mRNA is a versatile and promising tool for the delivery of transgenes into desired cells. https://mrna.creative-biolabs.com/custom-mrna-modification.htm
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
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.
5 Tips for Successful qRT-PCR Results InfographicQIAGEN
Market research shows that 66.6% of researchers use qRT-PCR for gene-expression profiling. Clearly, this is a very effective and popular technique for detecting RNA expression levels, but it’s still prone to pitfalls that can sometimes lead to disappointing results.
To help improve your experiments, we’ve made a new infographic with some interesting facts about qRT-PCR and 5 tips to help with your gene expression studies. This will cover everything from experimental design to data analysis.
Improving RNA yield and quality is easy with the right protocols. This trouble shooting guide presents methods to overcome common difficulties in RNA purification. Prevent RNA degradation, improve RNA yields, reduce genomic DNA contamination, and more.
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.
Multicopy reference assay (MRef) — a superior normalizer of sample input in D...QIAGEN
Copy number variations (CNVs) and alterations (CNAs) are a source of genetic diversity in humans and are often pathogenic. Numerous CNVs and CNAs are being identified with various genome analysis platforms, including array comparative genomic hybridization (aCGH), single nucleotide polymorphism (SNP) genotyping platforms, and next-generation sequencing. Independent verification of copy number changes is a critical step. Quantitative real-time PCR (qPCR) is a classic method to verify microarray copy number findings. Traditional copy number assays that use qPCR typically rely on a putative single-copy gene reference assay (e.g., RNase P or TERT) to normalize the DNA input for downstream ΔΔCT-based copy number calculation for comparison to a reference genome. When applied to cancer samples, these single-copy reference assays may no longer be a reliable indicator of DNA input due to the presence of complex chromosome composition (both in chromosome number and structure). To meet the need for an accurate DNA input normalizer, especially for heterogeneous tumor samples, QIAGEN developed a multicopy reference (MRef) assay for real-time PCR copy number analysis. This assay, in conjunction with QIAGEN’s greater than 10 million genomewide copy number assays and pathway- and disease-focused copy number PCR arrays (Figure 1), provides a successful solution for copy number analysis. This article will address the assay design considerations, development, and performance of this multicopy reference (MRef) assay.
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!
RT2 Profiler PCR Arrays: Pathway-focused Gene Expression Profiling with qRT-P...QIAGEN
This paper evaluates the performance of the newest technique for monitoring the expression of a panel of pathway- or disease-specific genes: the RT2 Profiler PCR Array System. The RT2 Profiler PCR Array System combines the quantitative performance of SYBR® Green real-time PCR with the multiple-gene profiling capabilities of a microarray.
The RT2 Profiler PCR Array is a 96- or 384-well plate containing RT2 qPCR Primer Assays for a set of 84 related genes, plus 5 housekeeping genes and 3 controls. The complete system includes an instrument-specific master mix and an optimized first strand synthesis kit. This paper presents experimental data showing that RT2 Profiler PCR Arrays have the sensitivity, reproducibility, and specificity expected from real-time PCR techniques. As a result, this technology brings focused gene expression profiling to any biological laboratory setting with a real-time PCR instrument.
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 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
PCR - From Setup to Cleanup: A Beginner`s Guide with Useful Tips and Tricks -...QIAGEN
This End-Point PCR Beginner´s Guide will not only give you a comprehensive overview of tools and techniques to help you to get the most out of your samples, but also give you information on dedicated solutions and complete workflows on multiplex PCR and PCR fragment analysis.
Course: Bioinformatics for Biomedical Research (2014).
Session: 2.3- Introduction to NGS Variant Calling Analysis.
Statistics and Bioinformatisc Unit (UEB) & High Technology Unit (UAT) from Vall d'Hebron Research Institute (www.vhir.org), Barcelona.
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.
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!
CD Genomics provides a fast, one-stop bacterial RNA sequencing solution from the quality control of sample to comprehensive data analysis. Please contact us for more information and a detailed quote.
A class of DNA sequencing techniques currently in active development is third-generation sequencing, commonly referred to as long-read sequencing. In comparison to second generation sequencing, also referred to as next generation sequencing, third generation sequencing technologies have the capacity to create noticeably longer reads.
The ability to easily and efficiently analyse RNA-sequencing data is a key strength of the Bioconductor project. Starting with counts summarised at the gene-level, a typical analysis involves pre-processing, exploratory data analysis, differential expression testing and pathway analysis with the results obtained informing future experiments and validation studies
https://www.shamra.sy/academia/show/5b06e01c54e75
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.
Learn from influencers. Influencers play a crucial role when it comes to marketing brands. ...
Use social media tools for research. ...
Use hashtag aggregators and analytics tools. ...
Know your hashtags. ...
Find a unique hashtag. ...
Use clear hashtags. ...
Keep It short and simple. ...
Make sure the hashtag is relevant.
De novo transcriptome assembly of solid sequencing data in cucumis melobioejjournal
As sequencing technologies progress, focus shifts towards solving bioinformatic challenges, of which sequence read assembly is the first task. In the present study, we have carried out a comparison of two assemblers (SeqMan and CLC) for transcriptome assembly, using a new dataset from Cucumis melo. Between two assemblers SeqMan generated an excess of small, redundant contigs where as CLC generated the least redundant assembly. Since different assemblers use different algorithms to build contigs, wefollowed the merging of assemblies by CAP3 and found that the merged assembly is better than individual assemblies and more consistent in the number and size of contigs. Combining the assemblies from different programs gave a more credible final product, and therefore this approach is recommended for quantitative
output.
DE NOVO TRANSCRIPTOME ASSEMBLY OF SOLID SEQUENCING DATA IN CUCUMIS MELObioejjournal
As sequencing technologies progress, focus shifts towards solving bioinformatic challenges, of which
sequence read assembly is the first task. In the present study, we have carried out a comparison of two
assemblers (SeqMan and CLC) for transcriptome assembly, using a new dataset from Cucumis melo.
Between two assemblers SeqMan generated an excess of small, redundant contigs where as CLC generated
the least redundant assembly. Since different assemblers use different algorithms to build contigs, we
followed the merging of assemblies by CAP3 and found that the merged assembly is better than individual
assemblies and more consistent in the number and size of contigs. Combining the assemblies from different
programs gave a more credible final product, and therefore this approach is recommended for quantitative
output
DE NOVO TRANSCRIPTOME ASSEMBLY OF SOLID SEQUENCING DATA IN CUCUMIS MELObioejjournal
As sequencing technologies progress, focus shifts towards solving bioinformatic challenges, of which sequence read assembly is the first task. In the present study, we have carried out a comparison of two assemblers (SeqMan and CLC) for transcriptome assembly, using a new dataset from Cucumis melo. Between two assemblers SeqMan generated an excess of small, redundant contigs where as CLC generated
the least redundant assembly. Since different assemblers use different algorithms to build contigs, we followed the merging of assemblies by CAP3 and found that the merged assembly is better than individual assemblies and more consistent in the number and size of contigs. Combining the assemblies from different programs gave a more credible final product, and therefore this approach is recommended for quantitative
output.
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.
Digital RNAseq Technology Introduction: Digital RNAseq Webinar Part 1QIAGEN
QIAseq RNA is a revolutionary turnkey solution for digital gene expression analysis by NGS. From 10 genes to 1000, from one sample to 100, QIAseq RNA delivers precise results on both ION and Illumina sequencing platforms. The data from QIAseq RNA is directly comparable to expression analysis derived from whole transcriptome sequencing or by qRTPCR, only better, cheaper, faster, and more flexible. This webinar will describe the principles of digital expression analysis by NGS, and review the features and benefits of the QIAseq system, options available, and the integrated data analysis package.
The Indian economy is classified into different sectors to simplify the analysis and understanding of economic activities. For Class 10, it's essential to grasp the sectors of the Indian economy, understand their characteristics, and recognize their importance. This guide will provide detailed notes on the Sectors of the Indian Economy Class 10, using specific long-tail keywords to enhance comprehension.
For more information, visit-www.vavaclasses.com
Model Attribute Check Company Auto PropertyCeline George
In Odoo, the multi-company feature allows you to manage multiple companies within a single Odoo database instance. Each company can have its own configurations while still sharing common resources such as products, customers, and suppliers.
The Roman Empire A Historical Colossus.pdfkaushalkr1407
The Roman Empire, a vast and enduring power, stands as one of history's most remarkable civilizations, leaving an indelible imprint on the world. It emerged from the Roman Republic, transitioning into an imperial powerhouse under the leadership of Augustus Caesar in 27 BCE. This transformation marked the beginning of an era defined by unprecedented territorial expansion, architectural marvels, and profound cultural influence.
The empire's roots lie in the city of Rome, founded, according to legend, by Romulus in 753 BCE. Over centuries, Rome evolved from a small settlement to a formidable republic, characterized by a complex political system with elected officials and checks on power. However, internal strife, class conflicts, and military ambitions paved the way for the end of the Republic. Julius Caesar’s dictatorship and subsequent assassination in 44 BCE created a power vacuum, leading to a civil war. Octavian, later Augustus, emerged victorious, heralding the Roman Empire’s birth.
Under Augustus, the empire experienced the Pax Romana, a 200-year period of relative peace and stability. Augustus reformed the military, established efficient administrative systems, and initiated grand construction projects. The empire's borders expanded, encompassing territories from Britain to Egypt and from Spain to the Euphrates. Roman legions, renowned for their discipline and engineering prowess, secured and maintained these vast territories, building roads, fortifications, and cities that facilitated control and integration.
The Roman Empire’s society was hierarchical, with a rigid class system. At the top were the patricians, wealthy elites who held significant political power. Below them were the plebeians, free citizens with limited political influence, and the vast numbers of slaves who formed the backbone of the economy. The family unit was central, governed by the paterfamilias, the male head who held absolute authority.
Culturally, the Romans were eclectic, absorbing and adapting elements from the civilizations they encountered, particularly the Greeks. Roman art, literature, and philosophy reflected this synthesis, creating a rich cultural tapestry. Latin, the Roman language, became the lingua franca of the Western world, influencing numerous modern languages.
Roman architecture and engineering achievements were monumental. They perfected the arch, vault, and dome, constructing enduring structures like the Colosseum, Pantheon, and aqueducts. These engineering marvels not only showcased Roman ingenuity but also served practical purposes, from public entertainment to water supply.
Instructions for Submissions thorugh G- Classroom.pptxJheel Barad
This presentation provides a briefing on how to upload submissions and documents in Google Classroom. It was prepared as part of an orientation for new Sainik School in-service teacher trainees. As a training officer, my goal is to ensure that you are comfortable and proficient with this essential tool for managing assignments and fostering student engagement.
We all have good and bad thoughts from time to time and situation to situation. We are bombarded daily with spiraling thoughts(both negative and positive) creating all-consuming feel , making us difficult to manage with associated suffering. Good thoughts are like our Mob Signal (Positive thought) amidst noise(negative thought) in the atmosphere. Negative thoughts like noise outweigh positive thoughts. These thoughts often create unwanted confusion, trouble, stress and frustration in our mind as well as chaos in our physical world. Negative thoughts are also known as “distorted thinking”.
Students, digital devices and success - Andreas Schleicher - 27 May 2024..pptxEduSkills OECD
Andreas Schleicher presents at the OECD webinar ‘Digital devices in schools: detrimental distraction or secret to success?’ on 27 May 2024. The presentation was based on findings from PISA 2022 results and the webinar helped launch the PISA in Focus ‘Managing screen time: How to protect and equip students against distraction’ https://www.oecd-ilibrary.org/education/managing-screen-time_7c225af4-en and the OECD Education Policy Perspective ‘Students, digital devices and success’ can be found here - https://oe.cd/il/5yV
The French Revolution, which began in 1789, was a period of radical social and political upheaval in France. It marked the decline of absolute monarchies, the rise of secular and democratic republics, and the eventual rise of Napoleon Bonaparte. This revolutionary period is crucial in understanding the transition from feudalism to modernity in Europe.
For more information, visit-www.vavaclasses.com
3. Next-generation sequencing is rapidly becoming the method of choice for
transcriptional profiling experiments.
In contrast to microarray technology, high throughput sequencing allows
identification of novel transcripts, does not require a sequenced genome
and circumvents background noise associated with fluorescence
quantification.
Furthermore, unlike hybridization-based detection, RNA-seq allows genome-
wide analysis of transcription at single nucleotide resolution, including
identification of alternative splicing events and post-transcriptional RNA
editing events.
Introduction
3
5. A typical RNA-seq experiment
Preparation of total RNA.
Depending on class of RNA to be sequenced (i.e. mRNA, lincRNA, microRNA etc),
enrichment is performed. Good quality total RNA is critical, although alternative
protocols for degraded RNA exist.
Library preparation.
Library preparation consists of:
RNA fragmentation. Unlike short RNAs, mRNAs are typically fragmented to smaller
pieces of RNA to enable sequencing.
Reverse transcription. First and second strand cDNA is reverse transcribed from
fragmented RNA using random hexamers or oligo(dT) primers.
Adapter ligation. The 5’ and/or 3’ ends of cDNA are repaired and adapters (containing
sequences to allow hybridization to a flow cell) are ligated.
Library cleanup and amplification. Libraries are enriched for correctly ligated cDNA
fragments and amplified by PCR to add any remaining sequencing primer sequences.
Library quantification, quality control and sequencing. Library concentration is
assessed using qRT-PCR and/or Bioanalyzer and is ready for sequencing.
Data analysis.
Downstream data analysis consists of quality control such as trimming of sequencing
adapters and removal of reads with poor quality scores followed by mapping reads,
analysis of differential expression, identification of novel transcripts and pathway
analysis.
Introduction
5
6. RNA-seq technologies
The three most widely used NGS platforms for RNA-seq are SOLiD and Ion Torrent,
both marketed by ThermoFisher, and Illumina’s HiSeq. All three platforms have similar
sample input requirements and sequences millions of cDNA fragments per run. Below,
sample preparation and pertinent application-specific advantages and disadvantages
are discussed.
Introduction
6
8. Ion Torrent and SOLiD libraries are both prepared using similar protocols.
Introduction
8
9. Non-coding RNA-seq
MicroRNAs are sequenced
by ligating RNA adapters to
each end of the mature
microRNA followed by
reverse transcription and
PCR (RT-PCR).
Introduction
9
12. The popularity of high-throughput next-generation sequencing
(NGS) ushered a new era in transcriptome analysis with RNA-seq.
A widespread application of RNAseq requires workflows tuned to
the sequencing technologies involved, sample types, desired
analysis as well as the availability of genomic and computational
resources.
Depending on the workflow used, the accuracy, speed, and cost
of analysis can vary significantly.
Thus, it is crucial to study the tradeoffs involved at different steps
of an RNA-seq analysis to get the best accuracy subject to the cost
and performance constraints.
Furthermore, figuring out the optimal workflow is even more
challenging since, in general, the best overall approaches may
have sub-optimal performance for a specific data set in terms of a
specific measure, which necessitates a comprehensive analysis of
workflows using a wide variety of data sets.
Introduction
12
13. They report the performance and propose a comprehensive
RNA-seq analysis protocol, named RNACocktail, along with a
computational pipeline achieving high accuracy.
Validation on different samples reveals that their proposed
protocol could help researchers extract more biologically
relevant predictions by broad analysis of the transcriptome.
Introduction
13
16. Several efforts have made to compare the performance of different RNA-seq analysis
tools.
However, these studies have mostly focused on a single RNA-seq analysis step, or their
workflow analyses were limited to one or two steps such as alignment and
quantification.
Thus, a comprehensive and systematic analysis of the RNA-seq data from different
perspectives can contribute significantly toward extraction of maximal insights from
RNA-seq data.
Research question
16
24. In conclusion, this a comprehensive assessment with detailed investigation
at each analysis step clearly outlines the current state of the RNA-seq
analysis.
This protocol highlights algorithm issues that warrant the attention of
researchers, leads to a broad-spectrum analysis protocol that can enable
researchers to unleash the full power of RNA-seq.
They envision that this approach will facilitate researchers in gaining better
and more comprehensive biological insights from their transcriptomic data,
as exemplified by the results of our pipeline, which is only one possible
instantiation of the comprehensive protocol.
Conclusion
24