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We narrowed to 255 results for: hal.2

Showing: 61 - 80 of 255 results

  1. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    Published
    April 18, 2024, 1:15 p.m.
    ... were depositing and requesting each year (Figure 2). We did a similar analysis with CRISPR pooled libraries...different categories (Figure 2C). Note that in Figure 2, the numbers on the X axis refer to distribution and...rather than a cumulative total.    Figure 2: Addgene CRISPR deposit and distribution by function...rapid adoption of new tools, illustrated in Figure 2, and the longevity of tools seen in Figure 3, indicates... the easy part; using them can be a bit more challenging. Addgene’s free educational resources help scientists...

  2. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    Published
    Jan. 10, 2017, 3:37 p.m.
    ...the absence of genome editing (Fig. 2 and 3) (5, 3). Figure 2: Gene activation with nuclease-competent...shown to exhibit more limited activity than SpCas9 (2). In many cases this is due both to lower relative...PMID: 24076762. PubMed Central PMCID: PMC3844869. 2. Chari, Raj, et al. "Unraveling CRISPR-Cas9 genome...different perturbations. Consider, for instance, the challenge of attempting to simultaneously and selectively...

  3. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ...select for optimal sybody candidates. Figure 2: Outline of the process for selection of sybodies ...identify and isolate sybodies targeting the SARS-CoV-2 Receptor Binding Domain found on the membrane-bound...Generation Toolbox kit. Find the Seeger lab's SARS-CoV-2 sybodies! References Geertsma ER (2013) FX Cloning...2020) Synthetic nanobodies targeting the SARS-CoV-2 receptor-binding domain. BioRxiv.  https://doi.org...access to routinely, and it can be especially challenging when trying to develop antibodies against unstable...

  4. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    Published
    Jan. 24, 2019, 2:24 p.m.
    ...Figure 2). We’ve also created constructs with containing chemical tags such as the SNAP-tag, Halo-tag, ...possible when each of the XFP genes is introduced at ~2 copies per cell per color, following a Poisson distribution...AAV titer - The XFP genes have to be introduced at ~2 copies per cell per color to achieve the highest color...

  5. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    Published
    May 2, 2019, 12:41 p.m.
    ...presented in a series of quick (10 minute) or very quick (2 minute) talks so we got to hear from everyone. Addgene...evolutionary tree at least 6-7 times and on at least 2 separate continents. Mimulus (Andrea Sweigart) Monkeyflowers...Organisms, Part 1 Five Popular Model Organisms, Part 2 ...share all of these new organisms continues to be challenging. Organism specific viral vector systems would...thus making progress on the NSF “G to P grand challenge.” Creating a productive, new scientific community...proteins, and DNA. Euprymna scolopes (Joshua Rosenthal) The Hawaiian bobtail squid has an extreme number... a different protein. Did you know that most cephalopods die after they lay their 1 clutch of eggs? Propagation...

  6. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    Published
    Sept. 7, 2017, 12:17 p.m.
    ...3 plates at different densities for selection). A 2% agarose solution is then melted (and thereby sterilized...96-well assay plate. We generally freeze at least 2 ampules each of 4 clones with different levels of ...of agarose, which is then overlaid with the firmer 2% agarose gel solution discussed above for picking....incubator, additional growth medium is added to about halfway up each well. Tips for using the agarose cloning...Time CRISPR User (by a 1st Time CRISPR User) The Challenges of Cell Culture Plasmids 101: Mammalian Vectors...

  7. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    Published
    July 31, 2018, 12:55 p.m.
    ...therapy had fallen - plummeted really -  from grace 2 years prior with the death of Jesse Gelsinger in an... the whole field to a grinding stop and only now, 2 decades later, is it finally back in the limelight...the other hand, have broad tropism in the eye. AAV-2, the first identified serotype and the one used in...reading 1. FDA approves gene therapy for blindness 2. Gene Therapy emerges from disgrace to be the next... completing her PhD and medical training in Ophthalmology. Gene therapy was in its infancy, but the eye...

  8. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    Published
    Oct. 18, 2018, 12:37 p.m.
    ...procedure to be performed using a simple two-step, 2 hr protocol prior to transformation (Figure 1). As...Primer design Using specific primer design (Figure 2), IVA cloning can be used to perform any plasmid modification... for each type of modification is shown in Figure 2, and is discussed in more detail below. The main requirement...Biotechniques 10.1 (1991): 62-66. PubMed PMID: 2003926. 2. Bubeck, Peter, Monica Winkler, and Wilfried Bautsch... to amplify away from the insertion site. Encode half the new sequence at the 5’ end of each primer, with...

  9. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    Published
    April 11, 2017, 2:30 p.m.
    ... to DFHBI in the presence of that metabolite (Fig.2). Spinach eventually evolved into Spinach2, a more...PMID: 25713381. PubMed Central PMCID: PMC4364232. 2. Baker, Monya. “RNA imaging in situ”. Nature Methods...emerging environmental pollutants.” Front Chem. 2014; 2: 41. PubMed PMID: 25019067. PubMed Central PMCID: ...biosensors.” TrAC Trends in Analytical Chemistry 27(2):108-117 · February 2008. 11. Svensen, Nina and Jaffrey...technology has been studied and used in assays for over half a century; the Addgene blog has an excellent primer...

  10. Predicting Adverse Reactions to Monoclonal Antibody Drugs

    Type
    Blog Post
    Published
    Feb. 13, 2024, 2:15 p.m.
    ...various components of the immune system. Figure 2: In-vitro models for immunogenicity prediction of ...drug reaction. This system would somewhat act like a 2-step verification. All approaches have their own limitations...Future Enhancements. Frontiers in drug discovery, 2, 952326. https://doi.org/10.3389/fddsv.2022.952326Joubert...Formation in Oncology: Clinical Relevance and Challenges. The oncologist, 21(10), 1260–1268. https://doi.org...

  11. Crowdfight, a Platform to Boost Scientific Collaboration During COVID-19 and Beyond

    Type
    Blog Post
    Published
    June 1, 2021, 1:15 p.m.
    ...mammal called alpaca could neutralize the SARS-CoV-2 virus. But while the early results were promising,...receiving dozens of requests every day.    Figure 2: When a researcher needs a collaborator for a specific...Crowdfight will: 1) Find an expert with the right skill; 2) Confirm the expert’s suitability and availability...lockdown forced us to put our research projects to a halt. None of us works directly with viruses or transmissible...

  12. Technical Design of a Western Blot

    Type
    Blog Post
    Published
    Aug. 22, 2024, 1:15 p.m.
    .... The most common options are 4 °C overnight or 1–2 hours at room temperature (RT). Most people choose...overnight at 4 °C and their secondary antibody for 1–2 hrs at room temperature, which allows them to reduce...Tris-glycine 6–400 kDa Tris-glycine 100 V, 1–2 hours Easy and cheap to handcast Short shelf-life...–40 kDa Tris-tricine 30 V, 1 hour or 100 V, 1–2 hours Good separation, quality, and stability ...separation of proteins of different sizes.  Table 2: Recommended gel percentages for various protein sizes...are consistent and stable at room temperature for 1–2 years and are available in formulas specific to your...response over a broad dynamic range that typically spans 2–5 orders of magnitude, which includes a wide dynamic...

  13. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    Published
    Jan. 16, 2020, 2:18 p.m.
    ... qPCR is that the results can vary by a factor of 2. This means that if you set up two identical assays...amplitude measured for each droplet in each well (Figure 2). A clean ddPCR should have a clear separation between...as low as 1C/s. Increasing the elongation time to 2 minutes and the denaturation time to 1 minute has ...generating the standard curve for qPCR can be challenging to get right. Here at Addgene, we use a linearized...droplet digital PCR for vaccine evaluation in challenged monkeys and TB diagnosis." Emerging microbes ...

  14. Plasmids 101: Protein tags

    Type
    Blog Post
    Published
    Dec. 11, 2014, 4:26 p.m.
    ...(you generally don’t want more tag than protein), 2-3 tags is increasingly common. Tandem affinity purification...TEV cleavage site (more on that in a moment), and 2 ProtA IgG-binding domains. TAP has since come to encompass...found in this paper (available at Addgene).   Table 2: Protease recognition sites commonly used with tags...purify proteins can face any number of technical challenges depending on their protein of interest. After...After several decades of trying to address these challenges, researchers have amassed a considerable molecular...

  15. Visualizing Translation at the Single Molecule Level

    Type
    Blog Post
    Published
    Aug. 1, 2017, 1:15 p.m.
    ...Find ribosome profiling plasmids at Addgene. 2. Halstead, James M., et al. “Translation. An RNA biosensor...the reporter mRNA is labeled by PCP-mCherry (Figure 2). The 3’ UTR also contains a CAAX sequence to bind...translation inhibitors cycloheximide and puromycin, Halstead et al. verified that translation is necessary ...translation is known to be important in development, and Halstead et al. used TRICK to examine translation of Drosophila...

  16. OCaMP: A New Calcium Indicator for Neural Imaging

    Type
    Blog Post
    Published
    Aug. 19, 2025, 1:15 p.m.
    ...the fluorescent protein mOrange2 (Figure 2). Figure 2: OCaMP structure and mechanism of action...module_attribute "schema_version" is_json="true" %}{% raw %}2{% endraw %}{% end_module_attribute %}{% module_attribute...developed a new calcium indicator, OCaMP. The challenge: Expanding the fluorescent toolbox Modern neuroscience...

  18. Antibodies 101: Avoiding the Mouse on Mouse Mess in IHC

    Type
    Blog Post
    Published
    Sept. 21, 2023, 1:15 p.m.
    ...recognize Fab fragments from the same species and 2) these fragments will be more likely to bind (and ...epitopes recognized by the secondary.   Figure 2: There are a few common solutions to avoid background...staining can be a challenge in the best of cases. But it can be particularly challenging when your only ...

  19. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    Published
    Oct. 1, 2015, 2:30 p.m.
    ... will repeatedly nose-poke at the active hole [1, 2].    Place preference/place aversion also models reinforcement...spend most of its times in the opposite chamber) [2]. Laser activation is usually controlled automatically...two different ways 1) your hypothesis is wrong or 2) there is some problem with the technical implementation...PMID: 21716290. Pubmed Central PMCID: PMC3775282. 2. McDevitt, Ross A., et al. "Serotonergic versus nonserotonergic...than ICSS. The test chamber is divided into two halves that are distinguished visually through different...

  20. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ... analogs such as 2-fluoroadenine (2-FA), organisms with functional APT will convert 2-FA to cytotoxic ...selection of plants undergoing RNAi, and LIIbeta F 1-2 RNAi, which allows assembly of intron-spliced hpRNAs...nucleotides, resulting in death. In the presence of 2-FA, potent gene silencing of APT is required for survival...such as in the model flowering plant Arabidopsis thaliana (Li et al., 2013; Zhang et al., 2018) and the ...
Showing: 61 - 80 of 255 results