NEB Scientific Posters
Your search returned 11 results.
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High-throughput methyl-binding domain-based DNA enrichment and sequencing
High-throughput methylated DNA enrichment can be an important tool to support high-throughput and large-scale epigenetic studies. Using a fused methyl-CpG binding domain from human MBD2 to the Fc tail of human IgG1 (MBD2-Fc) coupled with paramagnetic hydrophilic protein A beads, methylated DNA can be enriched from mixed genome samples and cell-free DNA (cfDNA) upstream of sequencing.
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Integration of whole exome sequencing and RNA-seq facilitates somatic mutation detection in tumor tissue samples
Somatic mutations are critical targets for disease diagnosis, prognosis, and drug discovery. Whole genome sequencing and targeted sequencing approaches are crucial for identifying somatic mutations in cancer. Meanwhile, RNA-seq has emerged as a complementary tool for somatic mutation profiling, especially for variants of uncertain significance (VUS), to characterize cancer types for precision medicine. Therefore, a comparative study to determine the concordance between DNA-seq and different RNA-seq methods and data analysis tools will help define the optimal approaches for somatic variant detection. This poster presents data from paired tumor and normal tissue samples from rectum and colon cancer patients, analyzing CNVs, SNVs, and indels.
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Accurate detection of small non-coding RNAs using NEBNext® Low-bias Small RNA Library Prep Kit
This poster describes a novel, ligation-based small RNA library preparation method that is characterized by reduced bias in addition to increased detection of sncRNAs. Libraries can be made in ~3.5 hours using a streamlined protocol with bead-based size-selections and cleanups. The robustness of this method is demonstrated with its compatibility across a broad input range (0.5 ng – 1,000 ng of Total RNA), as well as with challenging sample types, such as formalin-fixed paraffin-embedded (FFPE) RNA. The NEBNext Low-bias Small RNA Library Prep Kit sets a new standard for seeing small RNAs clearly.
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A sample-to-results workflow for Respiratory Syncytial Virus (RSV) whole-genome sequencing
RSV, a single-stranded RNA virus, is the leading cause of respiratory illness in infants. Using multiplex targeted amplification sequencing techniques, the presence of RSV in the population can be monitored for mutations that negatively impact treatment efficacy. Here, we describe a newly developed RSV sequencing approach based on amplicon-targeted sequencing for Illumina or Oxford Nanopore Technologies platforms, and featuring the NEBNext RSV Primer Module.
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Matched fresh frozen and FFPE patient tissues reveal the enhanced sensitivity and data quality of a novel DNA library prep method
Formalin-fixed, paraffin-embedded samples are a challenging sample type for most NGS library prep methods. We developed a novel method, compatible with both high and low quality FFPE DNA samples, employing three new enzyme mixes, designed specifically for compatibility with FFPE samples. The NEBNext UltraShear FFPE DNA Library Prep Kit includes an enzymatic fragmentation step that improves the library yield, library metrics, and variant calling accuracy.
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Streamlining RNA and DNA library construction methods to meet the challenges of high-throughput sequencing
With the goal of streamlining the processing of a variety of samples of various input amounts, the NEBNext UltraExpress® library prep kits respond to a user-stated need for faster, more efficient, and easily automated workflows. One key optimization is the single-condition workflow, which enables the simultaneous processing of multiple different sample input amounts (within the kit’s stated input range) with a single adaptor concentration and a single recommendation for PCR cycles. These advances have resulted in a single-tube solution, incorporating master-mixed reagents, reduced incubation times, fewer clean-up steps, and the generation of less plastic waste.
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E5hmC-seq: Detection of 5hmC at single-base resolution
DNA methylation is an epigenetic regulator of gene expression with important functions in development and diseases, such as cancer. Typically, the modified cytosines, 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC), are detected by sequencing Illumina libraries generated using a gentle, enzyme-based workflow called NEBNext® EM-seq, or by the harsher bisulfite conversion. Here, we describe an enzymatic method that enables specific detection of 5hmC, using the NEBNext Enzymatic 5hmC-seq (E5hmC-seq) Kit.
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Germline variant calling with EM-seq data
NEBNext Enzymatic Methyl-seq (EM-seq) workflows involve base conversion, which can be a challenge for variant detection. In EM-seq, this challenge is overcome using bioinformatic tools. Because methylation information is only preserved on a single strand in EM-seq libraries, the other strand can be used to detect genetic variation. Using this method, we can call germline SNPs with high precision.
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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.
