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We narrowed to 1,042 results for: URE

Showing: 201 - 220 of 1042 results
  1. Viral Vectors 101: Calcium Sensors

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    Blog Post
    ...noise and ensure you are measuring the events-of-interest. Sensors suitable for ER measurements include:... requiring only a simple measurement. However, single wavelength measurements are more susceptible to ... RE. Structure-and mechanism-guided design of single fluorescent protein-based biosensors. Nature chemical...subtype) and specific subcellular compartments for measurements. All of the sensors discussed below fall into...slower). GCaMPf may be suitable for very rapid measurements, like action potentials in neurons. jGCaMP7 ...additional versions are available to simultaneously measure calcium changes in the ER, cytosol and mitochondria...you combining your calcium reading with other measurements, specifically fluorescent ones? Many of the ...
  2. Corralling and Cornering the Goldilocks Antibody

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    Blog Post
    ...to compare exposure times of our samples. A prominent, strong signal has a shorter exposure time than ... be essential to your new research project, but unsure which one will mark your target the best, or even...The strength is based on binding specificity and exposure time. We first have a negative control which does...align with what the atlases (mentioned above) and literature suggest, providing an expectation and comparison...imaged pattern does NOT align with the atlases/literature and is unrelated to the target protein, we categorize... goldilocks antibody selection process The exposure time of the primary antibody compared to negative...control also determines the signal’s strength. Exposure time is the amount of time the sample needs to...
  3. Early Career Researcher Toolbox: Finding Relevant Papers

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    Blog Post
    ...analyzes publications and figures using machine learning to extract important features such as influential ... top of the ever expanding body of biomedical literature! PubMed PubMed is the classic search engine ...eventually replace the existing version of PubMed.  Key features This new version has an improved search algorithm...updated. This Twitter list contains >300 Twitter literature bots, including a bot for biosensor papers, microbe...Semantic Scholar Semantic Scholar is an AI-backed literature search developed by the Allen Institute for Artificial...that are more relevant to your search terms. Key features Semantic Scholar has several ways to focus your... populated at the top of this filter. A unique feature of Semantic Scholar search results are the topic...
  4. Cloning Mammalian Cells with the Agarose Method

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    Blog Post
    ...using cloning rings. A picture of a 60 mm dish ready for picking is shown in figure 1.   Find Additional... Protocol Pages Agarose cloning method procedure The procedure (illustrated in the YouTube video below...well plate is also prepared with about 200 ul of culture medium containing serum in each well. A 200 ul ...both Hepes and bicarbonate) is brought to room temperature and placed in the hood. The medium from a 10 ...is capped, swirled and tested for the correct temperature by placing on the wrist or cheek. Within less...warm but not hot- and still liquid- it is rapidly poured onto the middle of the plate, where it immediately...the agarose cloning method Test your agarose temperature. Cloning cells using an agarose overlay does ...
  5. CRISPR 101: Cytosine Transversion Editors

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    Blog Post
    ...editors for more details on how they work!) Figure 1: Base transitions are edits between the purine...side groups of DNA bases and leave the core ring structure intact, allowing them to make 4 out of the possible... Cas9 are sequence specificity and the R-loop structure. In order to bind DNA, Cas9 requires (1) a guide... a region of separate strands called an R-loop (Figure 2). This allows the guide RNA to base pair with...accessible on the outside of the enzyme. Figure 2: CRISPR R-loop. Created with BioRender.com. ...abasic sites with A's, producing C → A edits. Figure 3: Mechanism of cytosine transversion editors. ...experimented with adding different proteins to the CGBE architecture. They conducted a CRISPRi screen to identify...
  6. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

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    Blog Post
    ...kinetics which are illustrated in figures 2 and 3.  Figure 2: Glutamate titration curves for ...from Aggarwal et al…, Supplementary Figure 3.   Figure 3: pH titration curves and ∆F/F0 for...recommend using the SGZ for in-culture work only, GPI anchors for in-culture work and 2P in vivo imaging,...using both 1-photon and 2-photon imaging.   Figure 1: 1P and 2P excitation and emission spectra of...iGluSnFR3 v857. Image adapted from Aggarwal et al., Figure 1   Increased versatility through two different...Image adapted from Aggarwal et al…, Supplementary Figure 2.   Improved trafficking and localization... constructs. Image adapted from Aggarwal et al… Figure 4. PDGFR is a C-terminal fusion to the PDGFR transmembrane...
  7. 5 Great Apps for Lab Life

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    Blog Post
    ...content we store in them: music, emails, chats, pictures, and apps. Throughout my time as a graduate student...allowing you to upload anything from protocols to pictures of gels or even sketches to your notebook. The...saving trees! The Earth thanks you. 2. Gibco Cell Culture by Gibco Whenever I start to learn a new technique...I digress. When I started learning proper cell culture technique, I tried to find an app that could help...calculations. This is how I came across the Gibco Cell Culture app. Gibco organizes its app into four categories...are a lifesaver when your research requires cell culture and include a counter, lab timer, dilution calculator...calculator, and a couple of other nice features. This app also has a great educational section. This section...
  8. Neuronal labeling with Spaghetti Monster

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    Blog Post
    ...neuronal sub-cellular structures GFP is limited in its ability to label subcellular structures in the CNS. GFP...connections (Shepherd, 2004). Understanding the structure and function of these neural circuits is essential...experiments and visualize numerous molecules or structures at once (Shaner, 2007). In contrast to epitope...for the cells and affects the labeling of small structures like neurites. Like epitope tags, low FP expression...church of the Flying Spaghetti Monster; the smFP structure resembles images of the deity (see comparison ...processes. In contrast, smFPs can label fine neuronal structures with high fidelity and at lower concentrations...GFP. smFPs enable better labeling of these fine structures through their multiple, high-affinity binding...
  9. Hot Plasmids: Fall 2024

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    Blog Post
    ...solubility and stability (Figure 2).      Figure 2: Crystal structures of CRBNmidi alone (A) and...establishing CRBNmidi as a promising tool for future structure-guided drug design and studies of protein ...domains and is recommended for use in western blots (Figure 1). If you prefer to produce your own antibody ...chain are also available from Addgene!   Figure 1: Experimental validation for Anti-V5 [IPI-SV5...dimmer, showed greater photostability and also matured considerably faster, with expression levels comparable...comparable to GFP during early embryogenesis (Figure 3). mScarlet-I3 also offered clearer signals due to...autofluorescent tissues like the intestine. Figure 3: Expression of fluorophore-tagged GOLG-4 through...
  10. Twenty Years of Sharing: Addgene's Viral Vector Service

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    Blog Post
    ...growing in requests faster than jGCaMP7 (Figure 5B).   Figure 5: A) GCaMP6, jGCaMP7, and jGCaMP8 viral...setup. Teams across Addgene have worked hard to ensure that the service has grown steadily, helped by ...our catalog had grown to 879 viral catalog items (Figure 1). The service caught on quickly in the scientific...distributed over 100,000 viral vector preps!   Figure 1: A) Cumulative AAV preps available in the Addgene... they’re delivering their payload to.   Figure 2: Numbers of AAV tools, by category, available...respectively, by substrates, light, or engineered ligands (Figure 2). Biosensors, which include calcium and voltage...explain why they are the most commonly requested (Figure 3). Over 400 biosensors were distributed in 2023...
  11. Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research

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    ... (see Figure 1). Oligo 2: Add 'ggcc' to the 5′ end of your reverse complement guide (see Figure 1). Perform... tissue culture-free heritable genome editing using RNA virus-based delivery in wheat. Nature Plants. ...single guide RNA (sgRNA) used in Cas systems. The structure is also reminiscent of sgRNAs, with reRNA containing...gene-edited plants without the need for tissue culture (Maher et al., 2020; Qiao et al., 2025). However...choice of promoter for ωRNA (reRNA) expression (see Figure 1). Both plasmids contain a hygromycin resistance...phosphorylation and annealing of the two oligos. Figure 1: Components of TnpB, schematic of pKb-TnpB1 and...transformation into plant cells. Conclusion & Future Directions TnpB represents a new generation of ...
  12. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

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    Blog Post
    ... the process. Also refer to Figure 1 for a visual overview! Figure 1: General overview of the MoClo...dive directly into the pipetting! MoClo nomenclature and structure can be quite confusing for new users,...insert-to-vector ratio using the equation in Figure 3.    Figure 3: Calculation for the plasmid volume... MoClo a walk in the park.  Before you begin, be sure to check out Addgene’s MoClo guide for all the basics...assembly process to produce a multigene construct. Figure exemplifies the extraction and use of a coding ..., and have a clear vision of the final vector structure you want to achieve before starting your experiments...These overhangs are known as the common MoClo nomenclature. The overhangs are generated using type IIS ...
  13. AAV Titers: Where Do They Come From And What Do They Mean?

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    ...preparations are genome-containing (Figure 1).    Figure 1: Electron micrograph of an Addgene... by the temperature parameters. For example, we found that changing the annealing temperature of the PCR...viral vector solution, and generally these methods measure different characteristics of the solution. These... containing viral genomes. Physical titers are measured by quantifying the concentration of viral genomes... report the physical titer of a viral solution (Figure 2). Because physical titers are used for dosing...values mean and how they can be compared.   Figure 2: Label that comes on Addgene’s AAV aliquots. ...circled in red) report physical titer, which is measured by qPCR.   Physical titer is typically calculated...
  14. Deep Dive: Statistical Tests (Comparisons)

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    Blog Post
    ... of temperature on total crop production and the production window. A two-way MANOVA could measure the..., you could test the impact of temperature, humidity, sunlight exposure time, and watering volume on total...power. Not sure if your data meets the assumptions of parametric tests? If you aren’t sure if your variance...outliers or data spread within those data sets. Measures of dispersion – standard deviation/error, variance...in a certain direction (i.e., you just want to measure if a sample is greater than a control), then a ...variable. A two-way ANOVA could test the effect of temperature and humidity on an outcome such as total crop... one-way ANOVA can test either the effect of temperature or humidity on crop production.   MANOVA The ...
  15. Sequencing Options for CRISPR Genotyping

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    Blog Post
    ... Figure 1: Schematic representations of the indicated unbiased genotyping options. Figure reproduced...other elements, including those relating to DNA structure (e.g. epigenetic modification, bulges) that may...conventional algorithms may not provide a full picture of the impact of CRISPR editing in the model cell...negatives Combining sequencing techniques can ensure validated experiments Unbiased detection methods...sequencing and in silico prediction should give a broad picture of all possible editing events in the genome; from...induced by CRISPR-Cas nucleases in human cells." Nature biotechnology 31.9 (2013): 822-826. PubMed PMID...reveals RNA-programmed Cas9 nuclease specificity." Nature biotechnology 31.9 (2013): 839-843. PubMed PMID...
  16. Flow Cytometry Readouts: Yes, No, and Everything in Between

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    ...downregulation. Figure 3 shows a histogram for a yes-no dot plot, like the one shown in Figure 1.  ...populations in the histogram plot in Figure 4 compared to the dot plot in Figure 2. Quantification using MFI makes...Before you read out your plots, you’ll want to make sure you’ve also adjusted voltage and compensated your...adjusting voltage: During data acquisition, make sure that the signal for the positive population is no...should overlap with the negative control population (Figure 1). Based on where the GFP- population ends, you... the total cells present in the gate.   Figure 1: Example of yes-no flow plot of the (negative...expression, low expression, and everything in between (Figure 2). These smears tend to occur when you are looking...
  17. OCaMP: A New Calcium Indicator for Neural Imaging

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    Blog Post
    ...fluorescent protein mOrange2 (Figure 2). Figure 2: OCaMP structure and mechanism of action. Reproduced...brightness, ∆F/F0, or signal-to-noise ratio (Figure 3). Figure 3: One- and two-photon excitation and ... Figure 4: Calcium titration curves for OCaMP and various red and yellow GECIs measured with soluble...lasers available in your microscope setup. Figure 1: Excitation (dotted) and emission (solid) spectra...indicators spanning nearly the entire visible spectrum (Figure 1). The GCaMP series (e.g., jGCaMP8) remains the... yellow GECIs, OCaMP shows: Higher ΔF/F0 (the measure of signal change during calcium events), Greater...a CC-BY-NC-ND 4.0 International license. Figure 5: A) Simultaneous fluorescence (upper trace) and...
  18. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

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    ...Nishimasu H, Nureki O, Zhang F. Nature. 2015 Jan 29;517(7536):583-8. doi: 10.1038/nature14136. Epub 2014...Cas9’s crystal structure. “I thought maybe we could use information from the structure to guide and improve...technology. These libraries are lentiviral in nature, so please ensure that you are equipped and authorized to...genes, it is not possible to test each one and make sure it’s working. You can only do that screen with a... the SAM pooled library In a paper published in Nature in December, Konermann and her colleagues in Feng...-scale, knockout screening). As the Zhang lab's Nature paper reports, “Here we have shown that the SAM...Therapeutic uses in humans are also on the table. “In the future, there is potential there,” he says. "We have to...
  19. Plasmid Preps: Different Purity, Different Quantities, Different Uses

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    ...the purification procedure. Good indicators of DNA purity are the absorbance measurements at 230, 260, and...same. Before purifying a plasmid from a bacterial culture, it is important to consider your experiment.  ...Cloning grade DNA Transfection grade DNA Figure 1: Transformation introduces transformation grade...range). It can be extracted from small bacterial cultures (i.e. 2-3 ml) with or without commercially available...and DNA for transformation doesn’t need to be as pure as for other uses.   Cloning grade DNA Molecular...also dictate the required sizes of the bacterial cultures and commercial kits you will adopt for the extraction... cloning it is important that the preparation is pure from other nucleic acids (genomic DNA and RNA), ...
  20. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

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    Blog Post
    ...unique changes in their spectral properties upon exposure to a specific wavelength of light. PA-FPs can ...fluorescence, to a brighter fluorescent state by exposure to a specific wavelength of light. The switch ...switches from green to red fluorescence. mEosFP exposure to UV light causes an irreversible cleavage near...formation of mEosFP’s chromophore requires a temperature below 30°C and this is not ideal for experiments...emission of green fluorescence, but then further exposure to blue light also inactivates the chromophore...state. Dronpa can be reversibly reactivated by exposure to UV light. The fact that blue light both activates...photons are being emitted, making it easier to capture a clear image above background. The majority of...
Showing: 201 - 220 of 1042 results