This poster demonstrates the successful use of Monarch Mag Cell-free DNA for cfDNA extraction from liquid biopsy samples, leading to high-quality DNA library prep using a modified NEBNext Ultra II DNA library extraction protocol.
Automation of NGS library prep, enabled by robotic liquid handlers, meets the needs of NGS across a variety of utilities including basic research, drug discovery, and human Health. New England Biolabs and NEBNext® NGS solutions are designed with automation in mind by addressing several factors including fewer workflow components, streamlined workflows, reduction in bead cleanups, novel enzymatic solutions, and generous overages to support automation dead-volumes.
Streamlined NEBNext workflows empower researchers to efficiently generate high-quality sequencing libraries across diverse plant and animal sample types. By combining speed, sensitivity, and consistency, as well as compatibility with automation, they offer a powerful solution for accelerating genomic studies in agricultural research. NEBNext UltraExpress® DNA, NEBNext UltraExpress® FS DNA and NEBNext UltraExpress® RNA library prep kits provide rapid (2-3 hours) and reliable library construction, utilizing a single protocol for all input amounts (10 ng - 200 ng DNA and 25 ng - 250 ng RNA).
This attractive poster presents the range of applications for next-generation sequencing in the modern world.
Plasma cell-free DNA (cfDNA) analysis is advancing liquid biopsy for early disease detection, monitoring, and therapy selection, offering improved patient follow-up and cost efficiency over tissue biopsies. Technical challenges persist, including sample stabilization, low cfDNA concentrations, and extraction sensitivity. Monarch magnetic bead-based extraction enables efficient, reproducible isolation of high-quality cfDNA, recovering fragments as small as 50 bp and eluting in low volumes, ready for sequencing and digital PCR. Libraries prepared with NEBNext® Ultra II DNA Library Prep Kit show high conversion efficiency and sequencing quality, supporting sensitive variant detection. Duplex digital PCR assays using extracted cfDNA yield robust, accurate allele frequency quantitation for mutations such as PIK3CA p.E545K. The streamlined workflow integrates seamlessly with NEB downstream solutions, enabling reliable mutation detection from plasma samples and supporting multiple downstream applications.
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.
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.
We detail a workflow incorporating a magnetic particle processor, NEBExpress Ni-NTA magnetic beads, and NEBExpress Cell-free E. coli Protein Synthesis reactions that enables users to go from DNA to purified proteins in a single day with less hands-on time and greater reproducibility.
Single-chain antigen-binding fragments (e.g. VHH, scFv) offer several advantages over traditional therapeutic antibodies, including smaller size, enhanced solubility and thermal stability. Here we demonstrate the utility of our cell-free protein synthesis (CFPS) platform, NEBExpress® Cell-free E. coli Protein Synthesis (lysate), for the rapid production of antibody fragments in quantities suitable for direct analysis or further purification. CFPS workflows are fast, scalable and cost-efficient, and can be seamlessly integrated with our high-throughput DNA assembly methods to enable the rapid screening of diverse binder libraries. Additionally, the defined nature of these systems allows for straightforward optimization to improve expression of challenging constructs.
We demonstrate a simple workflow for universal in vitro DNA cleavage in cloning applications, called SpRYgest, using EnGen® sgRNA synthesis followed by in vitro cleavage with EnGen® SpRY Cas9 RNPs. We confirm that EnGen® SpRY Cas9 ribonucleoproteins (RNPs) can cleave more than 130 target sequences with various PAMs that are incompatible with wildtype Spy Cas9.
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