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TypeBlog Post...the time of manuscript submission. The rationale includes increasing the ability of reviewers (and, once...issue by requiring ever more condensed methods descriptions, legends and limiting numbers of figures such...could be mitigated by on-line services such as SlideShare, Figshare and Dataverse. Just as Addgene helps... raise the alpha isoform of GSK-3 to the same (deserved) stature as the beta isoform. Follow him on Twitter...
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Tips for Getting a Faculty Position
TypeBlog Post...Applying for a Faculty Position: the View from Both Sides," are summarized below. Check Out Erik Snapp's ...You want people to focus on you and your ideas. Slides will be distracting. What is the big question...feedback. Many of these documents are too vaguely described in other sources of guidance or the document examples...single malt whiskies, but I remember the days of modest postdoc salaries all too well. If there's a group... -
Five Popular Model Organisms, Part 2
TypeBlog Post...pathological developments (Vieira et al., 2019) Besides their ability to regenerate, the axolotl has several...scientific history, highly characterized genome, and widespread utilization, Arabidopsis has made significant...Arabidopsis genome project marked the beginning of widespread Arabidopsis genetics research development. Arabidopsis...allows for easy cloning of constructs specifically designed for use in Arabidopsis. There are also important... -
Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior
TypeBlog Post...same fluorescent markers, Brainbow 3.1 and 3.2 (described in the same publication) are unique because by...addition to the XFP in the construct, dBrainbow includes small epitopes fused to each XFP. Secondary antibodies...introduced using a plasmid construct called Zebrabow-GateDest containing an RFP, CFP, and YFP downstream of ...cell type. The Confetti mouse was first used to describe the self-renewal of cells in the intestinal crypt... -
Antibodies 101: Normalization and Loading Controls for Western Blots
TypeBlog Post...blot: we meet again. So useful, yet so finicky to design and optimize. Today we’ll cover the normalization...normalization can amplify other errors in experimental design and gel quantification. Figure 3: In a perfect...final blot. Quantifying the entire lane of protein provides a loading control that can be used for normalization...Antibodies 101: The Basics of Western Blotting Technical Design of a Western Blot Troubleshooting and Optimizing... -
Fluorescent Proteins 101: When GFP lets you down
TypeBlog Post...cofactor binding proteins as described above. The smallest tags are peptides that are engineered to bind...homology domains can be used to detect specific phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production...Péter, and Tamas Balla. "Live cell imaging of phosphoinositides with expressed inositide binding protein domains... -
Viral Vectors 101: Systemic Capsids
TypeBlog Post...expression to specific tissues and cell types. If desired, spatiotemporal control of transgene expression...systemic AAVs to find commonly used vector cargo designs and administration doses. Note: If capsid immunogenicity... Table 1 gives a brief overview of published descriptions of commonly used or emerging capsids. Selected..., B.E., Gradinaru V. Systemic AAV vectors for widespread and targeted gene delivery in rodents. Nature...Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain. Nature Biotechnology... prioritization for cell type-specific enhancer design. eLife,11, e69571. https://doi.org/10.7554/eLife... -
Technologies Enabled by NanoLuc® Luciferase
TypeBlog Post...above 600 nm. LumiFluors Schaub, F.X., et al. (3) designed two imaging probes. One consists of NLuc fused...Reporters. The LumiFluor reporter constructs were designed to be packaged into a retrovirus for infection...BRET probe for imaging. These authors set out to design a better fluorescent protein that could be co-imaged...acceptor, the more powerful the FRET signal. In well-designed FRET-based biosensors, the intensity of the FRET... -
Finding Your Perfect Job After University
TypeBlog Post...every item I tested was constantly tracked with barcodes and spreadsheets. It was great to be in the lab...industry positions. There were, however, some downsides to working in a large commercial company. In my...aspects of the company and gave me the freedom to design new projects and grow in the role. I understand... job different every day. I love the freedom to design and develop my own projects, but working remotely... -
When Fidelity Matters: A frank discussion about ligase fidelity
TypeBlog Post...end-joining, but will also efficiently ligate many undesirable structures, including substrates containing gaps...fidelity") better than T4 DNA Ligase (21, 25, 26). Despite this higher fidelity, Taq DNA Ligase can still ...possible NN pairs in the template. The probes were designed such that each possible pairing resulted in a ... high fidelity ligase, along with careful probe design and reaction condition optimization, will be critical... -
Quick Guide to All Things Lentivirus
TypeBlog Post...response element (RRE), an internal promoter, and the desired gene (transfer vector plasmid), 2) the HIV-1 genome...process is to test several protocols that have been described (protocol 1, protocol 2, protocol 3) to find the...quantify viral RNA. All of these methods have been described in a previous Addgene blog Once you know your ...Miller, A. DUSTY, and C. A. R. O. L. Buttimore. "Redesign of retrovirus packaging cell lines to avoid recombination... -
Finding Your Science Policy Path
TypeBlog Post...uncertainty of it all. Though I had spent the first few decades of my life following a very straightforward conveyor... harassment in academia or a large company who decides how they are going pursue regulatory approval for...their Local Engagement Playbook for step-by-step guides on how to engage in your local community. Read...comprehensive resource developed by Dr. Kelly Singel which provides an overview of all available science policy fellowships... -
Antibodies 101: Validation
TypeBlog Post...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...tuned for ready-to-use antibodies from Addgene Designing a recombinant validation experiment? Find plasmids... -
CRISPR Guide Update
TypeBlog Post...update, we have added new sections to share and describe some incredible new CRISPR technologies developed...much-anticipated prime editing section and additions describing CRISPR’s ability to make large genome edits using...transposases or the Cascade-Cas3 system. We also describe newly engineered Cas9s with more flexible requirements... -
Introducing the Addgene Antibodies 101 eBook!
TypeBlog Post...eBooks, the Antibodies 101 eBook is designed to be a helpful desktop reference. Each chapter brings together...an Antibody” quite useful. If you’re looking to design your first flow cytometry experiment, “Chapter ... -
Which Fluorescence Microscopy Technique is Best for Me?
TypeBlog Post...When performed correctly, fluorescence microscopy provides a high contrast image in which a bright signal...specificity to the viewer. Finally, modern microscope designs may further exploit the unique characteristics ...one object in the final image. Over the past two decades a number of techniques have been developed that...resolution and decreased phototoxicity. Lightsheet designs that allow for multi-view imaging can image thick... -
Antibodies 101: Introduction to Antibodies
TypeBlog Post...been infected with a microbe, to trigger immune cascades that clear these infections. Anything that generates...immune response and initiates different immune cascades. In the research setting, antibodies of different...antibody, which recognizes the primary antibody and provides the method of detection. You’ll also want to use...used for. Addgene’s Antibody Plasmid Collection includes plasmids that express antibodies, nanobodies, ... -
Countdown to Halloween @Addgene
TypeBlog Post...occurring earlier but that the photo evidence was destroyed to protect the innocent revelers. Our Halloween...for Halloween and how seriously we take costume design. In case you need more convincing, I’ve got just...to great customer support by dressing up as our Zendesk email system. Our 2013 winners dressed up as ... -
Performing In Vivo CRISPR Screens Using the FITS Approach
TypeBlog Post...single gene KO and pooled screening approaches; (4) encodes a Thy1.1 selectable marker to facilitate the isolation...assess T cell responses and clonal dynamics in lymph nodes (LN) and tumor cells. Figure used with permission...also reveal novel pathways. The FITS deposit also includes pLX_305_Ovalbumin, a plasmid that enables introduction... -
Penn Vector Core Partnership with Addgene
TypeCollection...Addgene partnership to provide AAV viral preps, includes listing of transfer status for viral preps....vectors See Addgene's Current AAV inventory , which includes all viral vectors transferred from the Penn Vector...