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We narrowed to 587 results for: des.1

Showing: 241 - 260 of 587 results

  1. Antibodies 101: Validation

    Type
    Blog Post
    Published
    March 24, 2022, 1:15 p.m.
    ...expected in another. You’ll need to validate that points 1 and 2 hold true under your specific experimental conditions...application and available resources.     Figure 1: Table summarizing the IWGAV five proposed antibody...like immunohistochemistry on human tissues, when (1) other strategies may not be feasible and (2) large-scale...validation: A view from the mountains. New Biotechnol 45:1–8. https://doi.org/10.1016/j.nbt.2018.08.002  Uhlen...to be present and recognizable by your antibody despite the manipulation. Additionally, not all sample ...large deletion that removes the entire exon that codes for the epitope recognized by your antibody. You...specific, then you expect that the bulk of the peptides identified to be from the target protein. This...

  2. Fluorescent Proteins 101: Visualizing Subcellular Structures & Organelles

    Type
    Blog Post
    Published
    June 22, 2017, 2:30 p.m.
    ...cell with the images from the Allen Cell Explorer (1). When studying the function of a protein or its role...various subcellular structures. Please see Figure 1 for commonly used markers for major mammalian organelles...Subcellular Localization Poster here! References 1. Allen Institute Plasmid Pages 2. Cooper GM. 2000....interest from its production site to its final destination. As an example, most secreted proteins are produced...pathway to their subcellular or extracellular destination. At the end of their life cycle, proteins may... about the subcellular structures your protein resides in and potentially which other proteins it forms...in Figure 2 (adapted from 4) - Dunn et al. 2011 provides an in-depth review of quantitative colocalization...

  3. RNA Extraction Without A Kit

    Type
    Blog Post
    Published
    March 24, 2020, 1:15 p.m.
    ...glassware should be treated with a DEPC solution for 1 hour, and autoclaved to remove residual DEPC. Alternatively...more large, expensive ultracentrifuge.  Figure 1: An outline of the different steps in RNA extraction...extraction: twenty-something years on. Nature Protocols 1:581–585. https://doi.org/10.1038/nprot.2006.83 Additional...Allewell and Sarma, 1974). Additionally, ThermoFisher provides a protocol on how to integrate RNAlater® with ...

  4. The Power Behind NGS Plasmid Validation: seqWell

    Type
    Blog Post
    Published
    April 19, 2017, 3:25 p.m.
    ...process is explained more fully in the animated Fig. 1 below. Normalization is a fundamental and often understated...plexWell™ libraries is well suited to large-scale de novo assemblies of short constructs like plasmids...all over the world. The rest of this piece will describe plexWell™ in more detail, and how we are using... containing fragments carrying sample-specific barcodes. Rather than continuing to process 100s to 1000s...

  5. Stem Cell Models for Disease & Open Science: Interview with Darrell Kotton

    Type
    Blog Post
    Published
    July 7, 2017, 1:32 p.m.
    ...Listen to the interview with Darrell Kotton Chapter 1: Introduction to Darrell Kotton and the Boston University...Unit, the Pulmonary Consultation Service, the Alpha-1 Center, and the Pulmonary outpatient clinic, all at...University Center for Regenerative Medicine. Darrell describes his career goals and what drives him as a praticing...

  6. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    Published
    Aug. 11, 2016, 2:23 p.m.
    ...some technical obstacles when working with virus: 1. Viral vectors undergo DNA rearrangements. Depending...approximately 20–50% of the packaging cells after 1–3 days of selection. A lower percentage of surviving...up-to-date tips for your virus experiments. References 1. Anthony, Simon J., et al. "A strategy to estimate... try amplifying the viral vector with bacteria designed to minimize rearrangements such as NEB Stable ...production of virus. Alternatively, if the virus encodes a fluorescent protein, the packaging cells could...working with, there’s a lot that goes into virus design, production, and transduction. Once you’ve harvested...

  7. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    Published
    Aug. 27, 2020, 1:15 p.m.
    ...tethered to bring them into close proximity (Figure 1). Formation of the complete FP results in a fluorescent...detected by fluorescence microscopy. Figure 1: How BiFC works. A split FP (e.g., yellow fluorescent... protein interactions in living cells. Nat Protoc 1:1278–1286. https://doi.org/10.1038/nprot.2006.201 ...This post was contributed by Patrick Miller-Rhodes from the University of Rochester Medical Center.  ...plasmids for your experiment at Addgene! How to Design a BiFC Experiment Much like FRET, BiFC experiments...quantification (Kerppola, 2006, 2013). These are described in more detail below: Fusion protein construction...each PIP and empirically test which combination provides the best fluorescent signal. As for linker sequences...

  8. Hot Plasmids: Summer 2025

    Type
    Blog Post
    Published
    Aug. 28, 2025, 1:15 p.m.
    ... high brightness and fast maturation time (Figure 1). See the paper for more details on small differences...mGold2t depending on the application. Figure 1: Super-resolution imaging of actin filaments with ...proteins for extended imaging. Nature Communications, 16(1), 3241. https://doi.org/10.1038/s41467-025-58223-5...., 2025). By co-delivering TRV1 with TRV2 that encodes ISYmu1 and an ωRNA under the same promoter, they...different applications require careful selection or design to ensure reliable readout and avoid biases. The...ex vivo HSC expansion cultures. A) Experimental design. B) UMAP of cell states in ex vivo expansion cultures...only at excitatory synapses; however, its modular design allows components to be swapped out, potentially...

  9. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    Published
    Sept. 8, 2016, 2:30 p.m.
    ...environments such as live cells and tissues (Figure 1). These flexible strategies give you the best of both...majority of in-cell labeling strategies have two parts: (1) a genetically encoded “tag” expressed as a fusion...Follow him on twitter @rhodamine110.   References 1. Liu, Zhe, Luke D. Lavis, and Eric Betzig. "Imaging...individual molecules. Poor photon budgets are a widespread issue with fluorescent proteins—even the thriftiest...student searching couch cushions for spare change, desperate to extract a few more photons from a sample. Chemical... the complexity of the system. A particularly desirable property for small-molecule labeling strategies...size, brightness, and cell permeability. In past decades, further refinements to this established dye structure...

  10. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...a rate of up to 1 per 500-1000 bp. In comparison, the DNA replication error rate is 1 per 109 bp. Unlike...common 30bp overlap sequences (Sites #1-5). Each overlap site encodes a unique restriction enzyme site and...Transcriptional Output-TANGO (PRESTO-TANGO) to screen the NCC-1 library of approved drugs against the entire kit; ...First, GMAP-compatible backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed. The authors...is multiplex automated genome engineering (MAGE) (1). MAGE uses recombineering (2) to simultaneously incorporate...of GFP (SEP) by a 9 amino-acid linker (see Figure 1). The key to the biosensor lies in pH: DsRed is relatively...inactivate SEP and leave only DsRed to fluoresce (1). With variants that can be targeted to specific cellular...

  11. Management for Scientists: Seeking Feedback

    Type
    Blog Post
    Published
    Jan. 8, 2015, 2:34 p.m.
    ...-public settings. Ask over coffee.  Have regular 1:1 meetings with team members (don’t become known for...communication between all members of a team. As described here in a 2012 Intuit blog post, a study coming...

  12. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    Published
    Sept. 23, 2014, 3:05 p.m.
    ...Recently I learned that Addgene’s pLKO.1 cloning protocol is viewed around 3,000 times each month. I ...on the Addgene website in December 2006, the pLKO.1-TRC cloning vector (deposited by David Root of the...would the thousands of other scientists using pLKO.1 variants learn about those updates? This is a problem...the site. Like Labguru, Quartzy offers software designed with scientists workflow in mind and can offer...

  13. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    Published
    June 4, 2020, 1:22 p.m.
    .... pH and DNases affect DNA stability Figure 1: So many choices! What to do? The main factors ...your downstream experiment. TE (10 mM Tris-HCl,1 mM EDTA, pH 8.0) buffer is the best buffer for preserving... favors the breakage of the hydrogen bonds, destabilizing the molecule and favoring the separation of ...nuclease free water is a great option because it provides a great versatility on how to use your plasmid...

  14. Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes

    Type
    Blog Post
    Published
    June 19, 2018, 1:38 p.m.
    ...optimized for expression in E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2). The key difference... plasmids are available from Addgene! References 1 Bourdeau, R.W., Lee-Gosselin, A., Lakshmanan, A., ...have hollow interiors and a protein shell that excludes water but is gas permeable. These vesicles are...sequentially exposed to acoustic pulses with high enough amplitudes to first collapse arg2 and then arg1 vesicles...

  15. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    Published
    Sept. 12, 2023, 1:15 p.m.
    ...while the paid version provides more tools for plasmid mapping and design (Figure 1). Benchling: While you...Davis from the University of Utah.   Figure 1: A plasmid map created in SnapGene of Blue Flame plasmid...primer design Design primers for Goldengate cloning   Design primers for In-Fusion Cloning Design primers...academic users. Serial Cloner: Free desktop-based software for plasmid design and mapping. ApE (A plasmid Editor...Primer design tools Many of the above-listed plasmid mapping tools have built-in primer design tools, ...stand-alone sites for general primer design, as well as primer design for different cloning applications...characteristics. NEB Data-optimized Assembly Design (DAD): This tool designs overhang sets for high complexity Golden...

  16. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    Published
    Feb. 14, 2018, 2:36 p.m.
    ...B. Gurumurthy, and Masato Ohtsuka. Nat Protoc. 13(1) (2018):195-215. PubMed PMID: 29266098 Creating knock-in..., with protocols using dsDNA templates achieving ~1-10% efficiency. Easi-CRISPR uses a long ssDNA template...working with CRISPR than just designing a repair template that contains your desired edit, and I want to make...say about CRISPOR, a tool to help you design, pick, and close guides for Cas9 and Cpf1. This tool supports...The blog post I mentioned, John Doench’s How to Design your gRNA for CRISPR Genome Editing, gets over ...these numbers show that this basic concept of gRNA design is still challenging for researchers using CRISPR..., and that standardizing and improving the gRNA design process and rules for various applications is necessary...

  17. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    Published
    June 9, 2016, 2:30 p.m.
    ...takes a different approach, randomly removing 1-20 nucleotides from the cleavage site specified by the gDNA...About NgAgo User's First Impressions Here References 1. Gao, Feng, Xiao Z. Shen, Feng Jiang, Yongqiang Wu...DNA guides NgAgo uses 5’ phosphorylated DNA guides (so called “gDNAs”) rather than the RNA guides employed...24 base DNA guides may be more user-friendly, since they can be ordered as oligonucleotides and should ...gDNA guides will also need to be transfected into the cells of interest. In contrast, RNA guides must ...important role in RNA interference, binding RNA guides to cleave foreign RNA. Argonaute proteins are also...cleave DNA in vitro, directed by phosphorylated DNA guides. Unfortunately, Swarts et al. were unable to show...

  18. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    Published
    July 27, 2021, 1:15 p.m.
    ...organoid model (Sun, et al. 2021).  Figure 1: Example schematic from the Organoid EasyTag system...without altering that of the internal control (Fig 1). Because this method relies on the fluorescence ratio...CRISPR inhibition (CRISPRi). A) Lentiviral vector design. B) Strategy for leak-free transcriptional control...Organoid EasyTag workflow combines thoughtfully designed plasmid components with Fluorescence Activated... heavy optimization. Claudia Vickers’s lab has designed novel SL fluorescent biosensors by integrating... rather than on the fluorescence intensity, it provides a direct and quantitative detection of SLs. The...biosensors could have a major economic impact.   Design and mechanism of action of the receptor-based SL...

  19. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    Published
    July 16, 2015, 2:30 p.m.
    ...microscopic resolution for studying colloidal gold (1).  The method was based on using a thin plane (sheet...Siedentopf, and R,. Zsigmondy.  (1902) Ann. Phys.  315, 1 – 39.
 A. H. Voie, D. H. Burns, F. A. Spelman. (1993...microscopy/en_de/products/imaging-systems/lightsheet-z-1.html ...After this initial application, Stelzer’s group described the single-plane or selective-plane illumination...and acquire images of a sample from two opposing sides and merge the images, such as in the Ultramicroscope...

  20. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    Published
    July 23, 2019, 12:46 p.m.
    ...determined LY6A, also known as stem cell antigen-1 (SCA-1), is the receptor used by PHP.B to reach the CNS...William L. Stanford. "Concise review: stem cell antigen‐1: expression, function, and enigma." Stem cells 25.6... a receptor for PHP.B. Read on to learn how the design of PHP.B may have contributed to this mouse-specific...Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain." Nature biotechnology...
Showing: 241 - 260 of 587 results