Join NEB’s Corporate Satellite Meeting on:
Advancements in NGS Sample Preparation
– Methylome Analysis, Targeted SNP Genotyping and Biobanking applications
New England Biolabs is a global leader in developing solutions for Next Generation Sequencing Sample Preparation and continues to push the forefront in providing high quality, robust products to support the clinical application of genomic data.
During this virtual „on demand“ satellite meeting, you will gain insights into the advantages of NEBNext NGS-library prep workflows in clinical applications:
- We will share first-hand wet-lab experiences of our users with you including recent successes in epigenetic analyses in acute or chronic kidney disease using cfDNA templates.
- You will hear about accurate sample identification and tracking applying in a biobank employing a novel high-throughput, cost-effective method for targeted SNP Genotyping.
- Moreover, we will share real-life experiences in how to best handle sequencing pipelines of up to 10.000 genomes per month.
Dr. Florian Erger, PostDoc, Inst. of Human Genetics, Cologne University Hospital, Cologne, Germany
Enzymatic cytosine conversion for epigenetic analyses of cell-free DNA
Cell-free DNA (cfDNA) has become an essential diagnostic tool, but its current uses like “liquid biopsies” or non-invasive prenatal testing mainly leverage genotypic differences and/or copy number variation…
Genotypic differences in the tissues-of-interest are generally only to be expected in the context of cancer, organ transplantation or pregnancy. For the majority of patients, these approaches are thus uninformative. Epigenetic profiling of cfDNA, on the other hand, promises almost universal applicability.
Bisulfite cytosine conversion has been the gold standard in DNA methylation analysis for decades. However, it causes significant DNA degradation, fragmentation and GC biases in the resulting sequencing libraries. As a proof-of-principle, we have performed combined cytosine methylation and nucleosome footprinting analyses on cell-free DNA using enzymatic cytosine conversion (NEBNext EM-Seq). We study 6 healthy control individuals and 6 patients with acute or chronic kidney disease. Direct comparisons with Bisulfite conversion are made. We show that enzymatic cytosine conversion significantly better preserves cfDNA fragmentation information and produces a more even sequence coverage. Compared to Bisulfite conversion, more CpG positions are covered at lower sequencing depths.
Whereas previously, separate experiments had to be performed to study cytosine methylation and nucleosome footprinting, reliable results for both can now be generated in a single assay. This represents a more comprehensive and inexpensive method for the epigenetic profiling of cfDNA, accelerating future discoveries in this fast-emerging field.
Andrew Barry, Sr. Manager Business Development at New England Biolabs, Ipswich, USA
A highly multiplexed target enrichment approach for sample identification and tracking using the NEBNext Direct Genotyping Solution
As next-generation sequencing is increasingly being adopted for applications including genetic screening and clinical diagnostics, preventing false reporting of results is imperative, thus demanding methods for positive sample tracking…
A reliable method to ensure sample provenance is to monitor single nucleotide polymorphisms (SNPs) that are highly discriminatory across individuals in an orthogonal assay that is performed upon sample accessioning, and comparison of genotypes with those obtained in a clinical assay. In order to incorporate a routine sample tracking method into diagnostic workflows, the method should be reliable, high-throughput, and cost-effective.
To address the need for targeted, high-throughput, sequencing-based genotyping assays, we developed the NEBNext Direct Genotyping Solution. This approach enables multiplexing of up to 96 samples in a single hybridization reaction that targets between 100 to 5000 SNPs. Here we demonstrate the power of this approach to distinguish unique human samples from each other using a sample identification panel of highly discriminatory SNP targets. Using this approach, minimal sequencing reads are required to obtain sufficient data for germline variant calling. With a single-day target enrichment and library preparation protocol and an approximately 12 hour sequencing strategy, data can be available within 24 hours of commencing sample processing. This approach presents a convenient and reliable method to ensure that data integrity is maintained in sequencing-based workflows.
Dr. Michael Quail, Principal Scientific Manager, Wellcome Sanger Institute, UK
Sequencing the UK Biobank – The nuts and bolts
Late breaking news: Updates on SARS-CoV-2 Sequencing included!
The Wellcome Sanger Institute in partnership with Decode Genetics is currently undertaking whole genome sequencing of the 500,000 UK Biobank cohort. Using NEB UltraII PCR free library prep we are preparing over 10,000 libraries per month. In this presentation I will give a background to the project, describe some of the challenges we have faced and the solutions we have implemented.
Your host is
Dr. Bjoern Textor, Senior Application Specialist, NEB Germany
Select products that assist Human Geneticists:
Learn more about reliable solutions from NEB
NEB offers a whole suite of reliable and high-quality research reagents and kits for Molecular Biology applications.
Here, we present popular NEB products Human Geneticists are typically using.
Simply click on the respective banner or Button to learn more and request a free sample!