Application Notes

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  High throughput silica membrane-based viral RNA extraction using positive pressure

  An application note about automated purification of viral RNA with Tecan's Resolvex A200 positive pressure workstation and MACHEREY-NAGEL's NucleoSpin 96 Virus Kit.

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  Restriction Endonucleases from NEB

  With over 40 years of offering restriction enzymes to the research community, NEB has earned the reputation of being a leader in enzyme technologies. Working continuously to be worthy of that distinction, NEB strives to develop enzymes of the highest purity and unparalleled performance.
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  Namocell's Two-Step Rare Cell Isolation

  Rare cell isolation can be very challenging using conventional cell isolation methods when the target cell population frequencies are less than 1% such as circulating tumor cells (CTCs) in blood or low transfected GFP-positive cells within a pool. Although cell sorting is typically performed to isolate rare cells, this is not ideal due to factors that may compromise the rare cell viability and recovery including long sorting time, high pressure (typically more than 20 psi), and large dead volume.
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  Ultra Low Sample Input with Namocell

  Effectively obtaining a single cell is key for single-cell transcriptomics, proteomics, and genomics. In this application note, we evaluate the single cell isolation efficiency of a 100 cells input with an adherent and non-adherent cell lines.
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  Namocell's SMART-Seq HT Protocol

  Single-cell cDNA Synthesis with Namocell Single Cell Dispenser and 1/4 reaction volume of Takara SMART-Seq® HT Kit.
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  A Practical Guide to Analyzing Nucleic Acid Concentration and Purity with Microvolume Spectrophotometers

  Microvolume spectrophotometers (MVS) are commonly used for the analysis of nucleic acid (NA) samples. They require a small sample volume (0.5–2.0 μl) and are economical, convenient and widely commercially available. Typically, they can measure NA concentrations as low as 1 ng/μl and they are compatible with various assays for all types of NA and proteins. Concentration and purity readings from MVS are often accepted as fact; however, a more detailed understanding of the readings can provide meaningful insight into the quality of the sample and its suitability for downstream applications. This technical note provides detailed explanations and guidance on how to interpret and use MVS measurements, with a focus on samples purified using silica spin columns.
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  Generating Clonal GFP Cell Lines

  Generating a clonal GFP cell line typically requires first sorting GFP positive cells with a cell sorter. Then GFP positive cells are cultured for several days to recover from the high pressure damage caused by the cell sorter. Clonal GFP cells are finally obtained by manual limited dilution at 1 cell per well. In this application note, Namocell Single Cell Dispenser was used to directly isolate single GFP positive cells into 96-well cell culture plate. Due to the low pressure of the Namocell Single Cell Dispenser, clonal GFP cell lines can be generated in a single step.
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  Single B Cell Genomics

  By use of a Namo Single Cell Dispenser, activated single B cells were isolated. Single B cell cDNA was suitable for both downstream single cell transcriptome library generation and PCR of the full length of immunoglobulin heavy and light chain variable domains.
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  Minimal Cell Input

  Obtaining a single cell is a critical step for single-cell transcriptomics, proteomics, and genomics. Traditional cell sorters have provided a powerful tool for sorting cells. However, it typically requires a large amount of cells. In this application note, Namocell Single Cell Dispenser was used to evaluate the single cell isolation efficiency of 100 minimal cell input with an adherent and non-adherent cell line. Complete your details and receive a copy of the application note in your inbox.
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  Single Nuclei Dispensing

  Using a Namocell Single Cell Dispenser, single nuclei were isolated and verified under a fluorescent microscope. Isolated single nuclei are suitable for single-nucleus RNA sequencing. Complete your details and receive a copy of the application note in your inbox.
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  NEBNext Direct® Custom Ready Panel technology analyzed using NextGENe®

  With targeted panels becoming an excellent alternative for whole exome and genome Next Generation Sequencing (NGS) there is an increasing need for highly flexible and optimized custom panels. The NEBNext Direct technology in conjunction with NextGENe software represents a high-quality solution for generation and analysis of sequencing data for identification sequence variants present in a user-friendly manner.

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  Mechanical DNA Fragmentation with the Q800R2 Sonicator

  The Qsonica Q800R2 Sonicator offers an alternative method for mechanical DNA fragmentation for Illumina TruSeq® Library Preparation Kits.

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  Restriction Enzymes for Droplet Digital PCR (ddPCR)

  NEB has evaluated a number of restriction enzymes for simple genomic DNA fragmentation in droplet digital PCR assays. This list is not meant to be exhaustive, but provides a starting point for validated enzymes with some simple usage guidelines. Each restriction enzyme listed below was successfully used for a ddPCR digest directly in the Bio-Rad® Droplet Digital SuperMix.

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  Analysis of chromosomal aberrations and recombination by allelic bias in RNA-Seq

  Read this publication in Nature Communications about the successful RNA purification for downstream RNA sequencing with the NucleoSpin RNA Plus Kit.
   

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  Characterize Immune Checkpoint Proteins and T Cell Exhaustion Using Multiplex IHC

  Advances in immuno-oncology have successfully led to novel cancer therapeutics with favorable patient responses that are more durable than conventional cytotoxic chemotherapy (1). However, not all patients respond to immunotherapy; therefore investigators are trying to identify clinically relevant biomarkers with the goal of developing therapeutics based on personalized medicine (2,3).

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  NucleoBond Endotoxin Free Plasmid DNA in CRISPR/Cas9 and lentiviral packaging/cell culture research

  Forward genetic screens are powerful tools for the unbiased discovery and functional characterization of specific genetic elements associated with a phenotype of interest. Recently, the RNA-guided endonuclease Cas9 from the microbial CRISPR (clustered regularly interspaced short palindromic repeats) immune system has been adapted for genome-scale screening by combining Cas9 with pooled guide RNA libraries. Here we describe a protocol for genome-scale knockout and transcriptional activation screening using the CRISPR-Cas9 system. 
  Read about the advantage of using NucleoBond Endotoxin Free Plasmid DNA from MACHEREY-NAGEL to avoid endotoxicity in virus production and mammalian cell culture. 

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  Hard-to-Lyse Samples

  Struggling with DNA isolation from hard-to-lyse samples? Up to 60% more DNA compared to standard extraction methods.
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  Chromatin / DNA Shearing with Qsonica Sonicators

  Qsonica offers several devices routinely used for shearing Chromatin and DNA. The Q800R system is our most advanced option and offers the highest throughput. In addition, the Q700 and Q125 are very popular and effective alternatives.

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  Nanoparticle Dispersion with Qsonica Sonicators

  Probe sonication is highly effective for processing nanomaterials (carbon nanotubes, graphene, inks, metal oxides, etc.). Sonicators have become the industry standard for different applications.

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  Cell Disruption and Homogenization with Qsonica Sonicators

  Cell lysis, tissue disruption and homogenization are common Sonicator applications. The ultrasonic energy output of each Sonicator model is adjustable and sonication parameters can be optimized according to your process requirements.

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  NEBuilder HiFi DNA Assembly versus Gibson Assembly

  This application note describes how to assembly multiple fragments and have 100% correct clones.

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  NEB Restriction Enzymes - Inside the Numbers

  After nearly 40 years of offering restriction enzymes to the research community, New England Biolabs (NEB) has earned the reputation of being a leader in enzyme technologies. NEB offers scientists 276 restriction enzymes, the largest selection in the industry.

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  Xtra Quick & Simple DNA Isolation

  A PhD student working at the Department of Immunology at the University Medical Center Utrecht shares her positive experience with the NucleoBond Xtra Midi Kit from MACHEREY-NAGEL

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  Cellular Imaging

  Visualization of Wnt receptor internalization by SNAP-tag technology from New England Biolabs (NEB)

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  Epigenetic landscape of proximal tubular epithelial cells using ChIPseq

  The epigenetic status of genes can be assayed by chromatin immunoprecipitation (ChIP). ChIP is a powerful technique to study DNA-protein interactions. To explore the genome-wide binding of transcription factors and the epigenetic landscape ChIP can be combined with sequencing (ChIPseq). This technique enables exploration of the epigenetic state in a global fashion

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  Gibson Assembly

  In this application note we describe the use of the Gibson Assembly™ Master Mix from New England BioLabs (NEB) together with gBlocks™ Gene Fragments from Integrated DNA Technologies (1) for the creation of a basic cloning vector for IP3R research.

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  How specific is your IHC antibody?

  Validating an antibody for immunohistochemistry (IHC) use involves more than just observing a signal in cells.

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  Q5 High Fidelity DNA Polymerase

  Marjorie Mercier, a scientist in Brussels, shares her positive experience with Q5 High-Fidelity DNA Polymerase for the amplification of the IgM variable region.

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  Q5 High Fidelity DNA Polymerase

  Colette Duez of the Center of Protein Engineering/ULg shares her positive experience with Q5 High-Fidelity DNA Polymerase for the amplification of a GC-rich (73%) insert of 1.4 kb from bacterial origin.

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  Q5 High Fidelity DNA Polymerase

  Mrs. Evy Vanderheyden, working at the VIB/KULeuven Molecular Microbiology and Biotechnology department shares her positive experience with Q5 High-Fidelity DNA Polymerase for the robust amplification of genomic yeast DNA.

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  Site-Directed Mutagenesis (SDM)

  Improved methods for site-directed mutagenesis using Gibson Assembly Master Mix from New England Bioloabs

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  Chromatin Immunoprecipitation

  SimpleChIP® in Literature: A synopsis of a recent publication by Tomas J. Bos, Ph.D., Department of Hematology and Immunology, Vrije Universiteit Brussel, Brussels, Belgium