NEB Scientific Posters
Your search returned 67 results.
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EM-seq™ enables accurate and robust methylation detection of cell free DNA and FFPE DNA sample types
NEBNext® Enzymatic Methyl-seq (EM-seq™) offers several improvements over traditional sodium bisulfite-based methylome analysis, owing – in large part – to the gentler, enzyme-based workflow. Less DNA damage enables longer reads with less sequencing depth. This poster summarizes the use of EM-seq with traditionally challenging sample types, cell-free DNA (cfDNA) and formalin-fixed, paraffin-embedded DNA (FFPE DNA).
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NEBNext® Ultra™ II FS DNA: A Robust Enzyme-based DNA Library Preparation Method Compatible with Plant Samples
Fragmentation is a bottleneck in the standard NGS workflow. The NEBNext® Ultra™ II FS DNA Library Prep Kit addresses this challenge with one-step enzymatic fragmentation, end-repair, and dA-tailing. Samples of plant tissue can be difficult to completely fragment without bias due to their molecular structure, but the Ultra II FS DNA kits enable robust library prep from Arabidopsis thaliana, Oryza sativa, and Zea mays samples.
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Enzymatic Methyl-seq: Next Generation Methylomes
DNA methylation is important for gene regulation. The ability to accurately identify 5-methylcytosine(5mC) and 5-hydroxymethylcytosine (5hmC) gives us greater insight into potential gene regulatory mechanisms. Bisulfite sequencing (BS) is traditionally used to detect methylated Cs, however, BS does have its drawbacks. DNA is commonly damaged and degraded by the chemical bisulfite reaction resulting in libraries that demonstrate high GC bias and are enriched for methylated regions. To overcome these limitations, we developed an enzymatic approach, NEBNext® Enzymatic Methyl-seq (EM-seq™), for methylation detection that minimizes DNA damage, resulting in longer fragments and minimal GC bias, here demonstrated with Arabidopsis thaliana and Cannabis sativa DNA.
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An E.coli Cell Lysate Based System for in vitro Protein Synthesis
The NEBExpress™ Cell-Free E. coli Protein Synthesis System is a high-performing, versatile and robust cell-free protein synthesis system developed by genetic engineering E. coli, optimizing a reaction buffer, and employing stringent manufacturing practices. This system was developed for coupled in vitro transcription and translation reactions for a variety of applications such as high throughput protein screening and engineering, as well as synthetic biology.
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An E.coli lysate-based system for in vitro Protein Synthesis
The NEBExpress™ Cell-Free E. coli Protein Synthesis System has been developed for coupled in vitro transcription and translation reactions resulting in high yields of proteins of various sizes (up to 230 kDa) and origins. A genetically engineered E.coli strain ensures stability of template DNA, RNA, and protein product. The Cell-free E.coli Protein Synthesis System is compatible with PURExpress Disulfide Bond Enhancer for better folding, and NEBExpress™GamS Nuclease Inhibitor for enhanced yield from linear templates. The reaction buffer formulation is compatible with SDS-PAGE (no acetone or TCA precipitation needed) and protein synthesis can be sustained for 10 hours at 37 °C or up to 24 hours at lower temperatures. Reproducible batches of lysate are produced, using highly stringent biomanufacturing processes and quality standards.
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Genome-wide profiling of nuclease protected domains reveals physical properties of chromatin (2019)
In metazoan cell nuclei, chromatin is functionally divided into transcriptionally active (euchromatin) or inactive (heterochromatin) regions. These heterochromatin regions constitute large chromatin domains that are in close contact with the nuclear lamina. Such lamina-associated domains (LADs) are thought to organize chromosomes inside the nucleus and are enriched for repressive histone modifications. Genome-wide profiling of heterochromatin, especially LADs, is often challenging and warrants a simpler and direct method. Here we developed a new method, Protect-seq, aimed at identifying regions of heterochromatin via resistance to nuclease degradation followed by next-generation sequencing. We performed Protect-seq on the human colon cancer cell line HCT-116 and observed overlap with previously curated LADs. We provide evidence that these protected regions are enriched for the repressive histone modification H3K9me3 and to a lesser extent H3K9me2 and H3K27me3. Moreover, the loss of H3K9me3 in human cells leads to an increase in chromatin accessibility. In sum, we demonstrate a novel technique to identify nuclease inaccessible regions of the genome and our data is consistent with the model that repressive chromatin domains are compacted and targeted to the nuclear lamina, likely via HP1 proteins, which act as scaffolds to maintain chromatin architecture.
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Genome filtering identifies species-specific DNA biomarkers for Mansonella perstans and Mansonella ozzardi, which enable differentiation of these closely related species and other co-endemic filarial parasites (2019)
Mansoneliasis is caused by infection with the parasites Mansonella perstans, M. ozzardi and M. streptocerca and is transmitted by insects such as biting midges and black flies. Immunosuppression caused by the parasitic infection may lead to worsening of other medical conditions. Mansoneliasis patients are often co-infected with multiple filarial parasites and anti-helminthic treatment is complicated. In this study, a bioinformatic filtering approach identified new diagnostic biomarkers, which were used to develop sensitive and species-specific LAMP assays that were validated on both patient and insect samples for point-of-care diagnostics.
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Draft genome sequences of Mansonella perstans and Mansonella ozzardi and their Wolbachia endosymbionts (2019)
Mansoneliasis is a widespread, yet neglected, filariasis of humans caused by infection with Mansonella perstans, M. ozzardi and M. streptocerca. Transmission to humans is via midge and black fly insect vectors whose endosymbiont is a member of a unique Wolbachia supergroup that is from both insect and filarial hosts. In this study, draft genome sequences of M. perstans, M. ozzardi and Wolbachia were obtained. This will provide insight into the biology and evolution of some of the most neglected filarial parasites.
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Choosing the Right Exonuclease (2019)
Download our poster to find the exonuclease best suited for your application.
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Enzymatic Methyl-seq: Next Generation Methylomes (2019)
DNA expression is tuned, both in nature and in the laboratory, with the application or removal of epigenetic marks, the most common of which being the methylation of cytosine residues. Historically, cytosine methylation at the single-base level has been detected by bisulfite sequencing, where sodium bisulfite is used to convert all unmethylated cytosines to uracils. This treatment is harsh, however, commonly leading to damage and even fragmentation of the very DNA meant to be sequenced; therefore, whole-genome bisulfite sequencing (WGBS) has substantial drawbacks.
Developed to address this challenge, NEBNext® Enzymatic Methyl-seq (EM-seq™) relies on a gentler, enzyme-based process for conversion of unmethylated cytosines, but not 5mC or 5hmC, to uracils. This poster introduces some of the ways in which EM-seq provides superior-quality sequencing metrics, including uniformity of coverage and detection sensitivity. For additional details, refer to the EM-seq product page.
