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  1. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ...reporter (Figure 2). The GUS enzyme cleaves a substrate, which will then form a blue precipitate. GUS ...Luciferases are great quantitative reporters and use a substrate to produce bioluminescence. Luciferases usually...fused to a protein of interest to elucidate the subcellular localization of your protein of interest. Find...
  2. New Tools Enable CRISPRa for Neuroscience Applications

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    Blog Post
    ...probe gene-function relationships in defined substructures of the CNS as well as individual neuronal populations...demonstrated by the Qi Lab, which developed a subgenome-scale CRISPRa screening approach in embryonic ...transduced with a pooled sgRNA library and were subsequently cultured for nearly two weeks to allow differentiation...
  3. Antibodies 101: Reading a Flow Plot

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    Blog Post
    ...license.    Pro tip! Subtypes of immune cells are often defined by a subset of the markers used to ...you’re interested in studying immunology or subpopulations of cells, you’ll soon find yourself encountering...
  4. Bioinformatics at Addgene

    Type
    Blog Post
    ...output of an initial process as the input to a subsequent process, we linked each step in our pipeline ... build one process at a time and connect each subsequent process to the existing ones. Once our first ...as output, which we specified as input to the subsequent bbmerge process.     Fig. 3: How we get...
  5. New CRISPR Tools: Cas7-11 and PASTE

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    Blog Post
    ... was composed of many protein subunits, it was nonetheless a new subtype. Cas7-11 had combined all the... complex Class I systems that contain multiple subunits, but the more they worked with it, the more they...
  6. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ... Cas13a binds and cleaves RNA rather than DNA substrates. In terms of structure, Cas13a shares no homology...Cas13a as a diagnostic tool, as discussed below. Subscribe to CRISPR Blog Posts Applications of Cas13a   ...hundreds of observed shRNA off-targets. They subsequently used catalytically inactive dLwaCas13a as a ...
  7. Data Freedom: The Expansion of Data Sharing in Research Publications

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    Blog Post
    ... data is to be made at the time of manuscript submission. The rationale includes increasing the ability...individual video. To PLOS’s credit, the organization subsequently clarified what the policy intent was and provided...be argued various authors have simply chosen to submit elsewhere). The majority of the scientific community...
  8. Harnessing Bacterial Toxins for Allelic Exchange

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    Blog Post
    ...counter-selection system relies on the sacB gene product of B. subtilis (Hmelo et al., 2015). The sacB gene product is...single crossover and integrated into the genome. Subsequent selection on sucrose will identify double crossovers...after about 48 hours, the resulting colonies turn subtly blue or magenta. This is particularly valuable ...
  9. Plasmids 101: Biotinylation

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    Blog Post
    ...a glycoprotein that consists of four identical subunits that can each bind one molecule of biotin. Biotin...surface labelling Attaching polypeptides to a substrate Flow cytometry/fluorescence-activated cell sorting...to attach proteins or polypeptides to a solid substrate (Holden and Cremer, 2003). References 1. Chapman-Smith...
  10. Filming Science Videos in the Age of Social Distancing

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    Blog Post
    ...around the lens. It allowed me to focus on the subject after I’ve left the room. Microphone: Lastly, I...shirt. But knowing I would have multiple interview subjects, I positioned a boom mic just out of frame allowing...camera from a distance if I needed to be close on a subject, or I could manually control the camera by hand...
  11. New Optogenetic Tools for Cytoskeleton and Membrane Control

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    Blog Post
    ...microtubule-destabilizing drugs are impossible to target to specific subcellular regions, laser ablation can damage other cellular...After 30 minutes of blue light illumination, substantial PA accumulates in the plasma membranes, indicated...reversibly control Rho GTPase activity on a global to subcellular scale, enabling precise control over vascular...
  12. Tips for Getting a Faculty Position

    Type
    Blog Post
    ... to Get a Faculty Position. This edition has substantial expansions on the chalk talks and Zoom interviews.... video interviews. For point of reference, I submitted my applications all by snail mail and only had... me and I'll try to answer your question and subsequently update the ebook. One of my graduate students...
  13. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

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    Blog Post
    ...published and unpublished experiments on similar subjects in a single, searchable space. Finally there are...2018). In this ideal situation, a researcher would submit an experimental design to a journal prior to performing...platform for pre-registration wherein researchers can submit designs and transparently reveal them upon completing...
  14. Antibodies 101: The Basics of Western Blotting

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    Blog Post
    ...wet transfer system the sandwich is completely submerged in a tank filled with a transfer buffer formulated...that emits light upon reacting with a specific substrate. The emitted light, or chemiluminescence, is detected...immunoprecipitation and are a common readout for subcellular fractionations, a procedure that isolates proteins...
  15. Technique: Probe Phage Genomes for Host Binding Proteins

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    Blog Post
    ...these fragments can be cloned into E. coli and subsequent proteins screened for cell-binding abilities ...proteins directly onto the nitrocellulose for subsequent probing for cell-binding activity.  First, cut...room temperature. After this, it is helpful to subject your membrane to several freeze/thaw cycles to ...
  16. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ...Fluorescence refers to the emission of light by a substance (fluorophore) that has absorbed light or electromagnetic...variant tag that covalently reacts with a small substrate ligand motif attached to a fluorophore. When paired...on a protein, enabling the labeling of protein substrates/peptides and antibodies. Example applications...
  17. Antibodies 101: Introduction to Antibodies

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    Blog Post
    ...antigen to myeloma cells. The hybridomas are then subcloned by limiting dilution, which produces an immortalized...based on molecular weight by SDS-PAGE, and are subsequently transferred to a membrane. A primary antibody...of antibodies is used to both identify the cell subsets of interest from among the mixture and to measure...
  18. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...is it? What parts of it move when you feed it substrate or add an essential cofactor? How many binding...especially since the potential rewards could be substantial: Plasmid tRNA Synthetase Expression Host ...Ermolenko, Dmitri N., et al. "Observation of intersubunit movement of the ribosome in solution using FRET...
Showing: 201 - 220 of 576 results