NEB Scientific Posters
Your search returned 82 results.
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Methods for sequencing cell-free DNA and cell-free RNA from human plasma
cfDNA and cfRNA are nucleic acid fragments found circulating in blood. These nucleic acids originate from various cell types, providing insights into the health of the system as a whole. By evaluating the change in cell-free nucleic acids over time, via serial blood draws, researchers can monitor for atypical biological processes, including cancer. When combined with the new Monarch Mag Viral DNA/RNA Extraction Kit, both the NEBNext Ultra II and the NEBNext UltraExpress library prep kits produce high-quality libraries.
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An end-to-end enzymatic solution for methylomes from low-input DNA
The cytosine modifications 5mC and 5hmC are important regulatory marks within the genome, influencing gene expression. Older techniques of methylome analysis, like bisulfite sequencing, are harsh and can damage the input DNA, decreasing the quality of the resulting libraries. NEBNext Enzymatic Methyl-seq (EM-seq) was introduced in 2019, and brought to bear gentle, enzymatic fragmentation, minimizing damage to the DNA and improving the quality of the resulting libraries. As a further improvement on the EM-seq workflow, NEBNext Enzymatic Methyl-seq v2 has been introduced, requiring lower DNA inputs and offering improved compatibility with lower-quality samples.
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Streamlined whole-genome and targeted sequencing workflows for prevalent RNA viruses: SARS-CoV-2, Respiratory Syncytial Virus, and Influenza A
Monitoring respiratory viruses through whole genome and targeted sequencing is now more important than ever before, particularly in the wake of the global pandemic. We have developed sequencing approaches for respiratory RNA viruses, including SARS-CoV-2, RSV, and Flu, to support scientists and public health laboratories to monitor these critical pathogens.
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Application of High Complexity Golden Gate Assembly to Rapid Engineering of Bacteriophage Genomes
Data-optimized Assembly Design (DAD) enhances Golden Gate Assembly (GGA) by using precise ligation fidelity measurements to select high-accuracy fusion sites. This approach replaces traditional GGA methods, enabling the efficient assembly of multiple parts while minimizing errors. We have developed a set of tools to apply DAD to the design of complex assemblies of up to 36 parts and 50kB final size in a single reaction. Combined with optimized reagents, DAD yields sequence-accurate constructs ready for use without further processing. The small parts are easy to produce via PCR or DNA synthesis and can be manipulated in E. coli plasmid systems, supporting viral mutagenesis and gene swaps. These principles facilitate modular assembly of bacteriophage genomes for high-throughput pathogen research.
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Utilizing Lyophilized LAMP Reagents in Rapid Molecular Assays
Loop-mediated isothermal amplification (LAMP) has become a widely used method for detecting target nucleic acids (DNA and RNA) as it offers a robust and simple alternative to PCR. LAMP is particularly well suited to point-of-care (POC) applications such as COVID-19 diagnostics because only a single incubation temperature is required for nucleic acid amplification, and the technology is compatible with simple detection strategies, including colorimetric or lateral flow readouts. To further increase the utility of LAMP in POC molecular diagnostic workflows, eliminating cold chain requirements for reagent shipment and storage is desired and can often be accomplished by lyophilization. This study investigates single and multiplex detection of several viral infectious diseases using lyophilized LAMP/RT-LAMP reagents.
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NEBNext UltraExpress™ RNA: A fast and flexible workflow for stranded RNA-seq library preparation
As the use and throughput of RNA sequencing continues to increase, there is a growing need for faster, more streamlined workflows that generate high-quality libraries. We have developed a kit with a 3-hour library prep protocol that enables the creation of high-quality directional RNA libraries in a single day when paired with poly(A) mRNA enrichment or rRNA depletion kits. The NEBNext UltraExpress RNA Library Prep Kit uses a single adaptor concentration and PCR cycle number for all RNA inputs (25 - 250 ng total RNA) and incorporates master mixed reagents, reduced incubation times and fewer cleanup steps, thereby reducing the total time and consumables used.
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NEBNext® solutions for challenging methods and samples
NEBNext UltraShear® and NEBNext UltraShear FFPE DNA Library Prep Kit excel in preparing DNA libraries, especially from complex samples like formalin-fixed, paraffin-embedded (FFPE) DNA. These solutions effectively fragment genomic DNA, enhancing library yields and sequencing metrics. Their use, particularly with the NEBNext FFPE DNA Repair v2 Mix, improves yield and coverage. Comparative studies with Covaris® ME220 demonstrate NEBNext UltraShear’s comparable methylation detection but with higher yields. The kit significantly reduces unmapped, chimeric, and foldback reads, lowering artificial mutation frequencies, and outperforms other kits in hybrid capture libraries for on-target coverage.
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E5hmC-seq™: Detection of 5hmC at single base resolution
DNA methylation, crucial in development and diseases like cancer, involves epigenetic regulation through 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC). Traditional detection methods, such as NEBNext® EM-seq™, cannot differentiate between these two cytosine forms. The new NEBNext Enzymatic 5hmC-seq (E5hmC-seq™) method overcomes this limitation by specifically identifying 5hmC. It glucosylates 5hmC to prevent deamination, allowing clear discrimination from 5mC and cytosine. Tested on human brain DNA, E5hmC-seq shows accurate 5hmC measurement with minimal GC bias, maintaining data quality. This advancement in identifying 5hmC is pivotal for understanding its role in cellular processes and diseases.
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Streamlined, Express DNA Library Preparation Methods Meet Requirements of High-throughput Library Construction
The importance of DNA sequencing as a laboratory technique continues to grow, and with it, the importance of faster, more streamlined workflows that generate high-quality libraries. The NEBNext UltraExpress DNA Library Prep Kit, for pre-sheared DNA and the NEBNext UltraExpress FS DNA Library Prep Kit, for intact DNA, take you from sample to library in under two hours. The streamlined workflow uses a single adaptor concentration and PCR cycle number for all inputs (10-200 ng) and incorporates master mixed reagents, reduced incubation times and fewer cleanup steps.
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Genomes of the Wolbachia endosymbionts from the human filarial parasites Mansonella perstans and Mansonella ozzardi reveal multiple origins of nematode-Wolbachia symbiosis
Wolbachia are Gram-negative, obligate intracellular bacteria in some filarial nematodes and about 60% of arthropods. The filarial parasites Mansonella perstans and Mansonella ozzardi harbor Wolbachia. Wolbachia have been consistently detected in M. ozzardi, but in M. perstans, the presence of Wolbachia may be isolate-dependent. Phylogenetically, the majority of Wolbachia from filarial parasites cluster into supergroups C, D and J, while arthropod Wolbachia are clustered in supergroups A, B, E, H and S. Wolbachia from Mansonella (wMpe, wMoz) are different from other filarial Wolbachia as they are placed in supergroup F, with Wolbachia from insects such as the bed bug. We present here the genomes of wMpe and wMoz, representing the first genomes from filarial Wolbachia of supergroup F. We also present two new genomes of arthropod Wolbachia from supergroup F.
