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We narrowed to 618 results for: abo.2

Showing: 221 - 240 of 618 results
  1. CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

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    Blog Post
    ...initial work was with TALENs, in collaboration with the Porteus lab [2]. With the advent of CRISPR/Cas9...nucleases. Genome Res. 2010 Jan;20(1):81-89. PubMed. 2. Bauer DE, Kamran SC, Lessard S, Xu J, Fujiwara Y,...PhD student at Harvard Medical School in the laboratories of Stuart Orkin and Daniel Bauer. He received...
  2. Production of Virus in Insect Versus Mammalian Cells

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    Blog Post
    ...) 38 kb Cells HEK-293T Sf9 Time to collection 2–4 days 5–10 days Purification Ultracentrifugation...Centrifugation Titration time ~1 day (AAV); ~1 week (lenti) 2–10 days Initial titers Higher Lower Resources... a factory to produce adeno-associated virus type 2 vectors. Human Gene Therapy, 13(16), 1935–43. https... widely available, so you’ll only have to worry about cloning your transfer plasmid. The transfer plasmids...typical insect cell preparations, which we talk about below. Processing The viral supernatant is typically...available, so the amount of cloning required is about the same between insect and mammalian systems. Baculovirus...dead cells and cellular debris. While adding time (about three to four days), this second round provides ...
  3. Plasmids 101: Control Plasmids

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    Blog Post
    ...the result in Figure 1 could look like this: Figure 2: Expression of Plasmid B (as internal control) This...down expression of Gene X but, as shown in Figure 2, this was likely due to the original transfection ...PMID: 20074371. PubMed Central PMCID: PMC2817684. 2. Replicates and repeats—what is the difference and...optimization within your experiment. Read on to learn more about the various controls that can be used for plasmid-based...: Expression Level of Gene X The results shown above are from a single experiment wherein Plasmid A (...be compared. The Empty Vector Control (mentioned above) could also serve as an important negative control...experimental cells. For example, in the experiment above, Plasmid A contains an shRNA that targets human ...
  4. Droplet Digital PCR for AAV Quantitation

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    Blog Post
    ... 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 ... droplet generator, you will have to be careful about transferring your droplets from the droplet generator...Additional resources on the Addgene blog Read more about polymerase chain reaction Browse our blog posts ...collection of viral vector related protocols Learn more about the Addgene viral service ...
  5. Tips for arabidopsis transformation

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    Blog Post
    ... Plant Methods. BioMed Central, 2(1), p. 19. doi: 10.1186/1746-4811-2-19. PubMed PMID: 17087829. PubMed...PMID: 24032037. PubMed Central PMCID: PMC3759027. 2 Harrison, S. J. et al. (2006) ‘A rapid and robust ... J. (2002) Arabidopsis : a laboratory manual. Cold Spring Harbor Laboratory Press. Available at: https...https://books.google.at/books/about/Arabidopsis.html?id=IfZAMNPWVk4C&redir_esc=y (Accessed: 21 September ...
  6. Neuronal labeling with Spaghetti Monster

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    Blog Post
    ...experiment using smFPs and standard neuronal tracers. 2. Visualization of neuronal sub-cellular structures...similar proteins. For example, N-cadherin (cadherin-2) is a postsynaptic cell adhesion protein that plays...PMID: 25915120. PubMed Central PMCID: PMC4573404. 2. Shepherd, G.M. (2004). In "The synaptic organization...enable accurate labeling of neurons. As discussed above, weakly expressed proteins in particular are difficult...resembles images of the deity (see comparison image above). The church of the Flying Spaghetti Monster is ...tags. 4. High-resolutions microscopy As indicated above, smFPs can be used with conventional microscopes...
  7. Hot Plasmids: Summer 2025

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    Blog Post
    ...synaptobrevin 2; At = ALFA-tag; BACE = β-secretase; AF = AMPA.FingR; FingR = fibronectin intrabody generated...RNA guide (known as an omega RNA, or ωRNA) (Figure 2), but TnpB is small enough to be carried by viral ...than a third of the size of Cas9). Figure 2: Comparison of TnpB (A) and Cas9 (B) genome editors...Check out our blog post on AAV serotypes for more about the tissue tropism of these and other AAVs. ...
  8. The AAVantages of AAV in CRISPR Screens

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    Blog Post
    ...handled at BSL-1, while lentivirus requires a BSL-2 setup. This requires a less involved setup when working...infection rates in your experiment. Figure 2: Visual representation of common AAV serotypes used..., 10(1). https://doi.org/10.1038/s41467-019-12449-2 Pupo, A., Fernández, A., Low, S. H., François, A.,...Hanlon, K. S., Kleinstiver, B. P., Garcia, S. P., Zaborowski, M. P., Volak, A., Spirig, S. E., Muller, A.,...vivoscreening platform. bioRxiv (Cold Spring Harbor Laboratory). https://doi.org/10.1101/2023.06.13.544831 Santinha...
  9. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

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    Blog Post
    ...properly exiting the ER and fluorescing (see Figure 2) [5].  When using FPs containing two cysteine residues...compartments including the secretory pathway (Figure 2), the inner membrane space of mitochondria and chloroplasts...PMID: 18228466. Pubmed Central PMCID: PMC2875081. 2. Snapp, Erik Lee. "Fluorescent proteins: a cell biologist's...moxFPs over standard FPs for the reasons listed above. In addition, because the moxFPs are highly monomeric...Medicine. Her doctoral research was conducted in the laboratory of Erik Lee Snapp and focused on optimizing fluorescent...
  10. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

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    Blog Post
    ...upstream of an opsin gene (e.g., channelrhodopsin-2, ChR2) fused to a reporter gene (e.g., mCherry) to...the correct orientation to be transcribed (Figure 2) (Sharma and Zhu, 2014). How does FLEx enable strong... Abdallah, Khaled, et al. "Adeno-associated virus 2/9 delivery of Cre recombinase in mouse primary afferents...Deniz, et al. "A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range ...FLEx enters the scene. FLEx: A light when night is about you! The FLEx (for flip-excision) switch (aka DIO...PMC4166436. Resources on the Addgene blog Learn more about FLEx vectors Download our Plasmids 101 eBook Read...
  11. Addgene-Seeding Labs Plasmid Grant: Accelerating Science Globally - 5 Scientists, 4 Countries & 3 Continents!

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    Blog Post
    ...the reaction’s metabolites. One useful genetically encoded H2O2 probe is pQE-30-HyPer3 (2) which he will...Pharmacology 2017 Jun 15;134:1-17. PubMed PMID: 27984002. 2. Bilan, Dmitry S. et al. “HyPer-3: A Genetically Encoded...The Addgene-Seeding Labs plasmid grant is a collaborative endeavor providing plasmids to researchers in... of Zimbabwe, Zimbabwe Dr. Idah Sithole-Niang (above, left) is an Full Professor in Biochemistry with... CRISPR effect observed in bacteria. Learn more about the different CRISPR applications in plants.     ...Rodrigo Arturo Rivera Solís, Associate Professor (above, far right), thrives to make affordable biosensors... to the Addgene blog for updates and future collaborations. Many thanks to all of the awardees, all who...
  12. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

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    Blog Post
    ...the team (Fig. 2). These images are based on the concept of bacteriographs created about a decade ago. ...spatially precise the activation is.   Figure 2: A bacteriograph of Michelangelo's "The Creation of...Twitter, you might have seen the video of her singing about the paper recently published by her lab and Mustafa...professor at the University of Freiburg. This paper is about a new tool called BLADE (blue light-inducible AraC....   Inspired by @UriAlonWeizmann, here my song about our paper recently published in @nchembio Title:...Additional resources on the Addgene blog Learn more about inducible promoters Visit the Plasmids topic overview...
  13. Summer SciComm: Science Storytelling

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    Blog Post
    ... (1) the struggle between two opposing forces or (2) the result of competing desires or (3) the presence... an introduction, and a resolution.   Fig. 2: The structure of a story - not just for fiction! ... your science with others.    My favorite quote about science ever comes from an interview in which someone...storytelling business like to say, science is all about navigating conflict. In literary terms, conflict...find that once I sit down and talk to a scientist about their research, these conflicts come out quite naturally... a simple question, like “Have you ever thought about how X works?”, or even by introducing a person that...devices, we sometimes forget the most important thing about science: It’s cool. Like bowties! Using science ...
  14. Hot Plasmids: Winter 2025

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    Blog Post
    ...resistance to the herbicide chlorsulfuron (Homo line 2) or wild type (WT), two weeks after herbicide was ... CLIPf tag and a C-terminal SNAPf tag; see Figure 2), the authors developed a cost-effective expression... when associated with microtubules. Figure 2: CLIP-SNAP fusion proteins for single-molecule FRET...synaptic events By Brian O’Neill The Lin Lab and collaborators recently created ASAP5, a faster and more responsive...
  15. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

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    Blog Post
    ... flanking sequences to their input string (Figure 2). This string can then be used as input for the prediction...top 5) and test them in their labs. Figure 2: Guidelines to predict pegRNA efficiencies with pridict.it...Biotechnology. https://doi.org/10.1038/s41587-024-02268-2 Schep, R., Brinkman, E. K., Leemans, C., Vergara, ... al., 2023). We trained our models in close collaboration with the Krauthammer lab (University of Zurich...
  16. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

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    Blog Post
    ...Epub 2016 May 2. PubMed PMID: 27136078. Find plasmids from this publication at Addgene. 2. Swarts, Daan...impaired NgAgo activity, with three mismatches abolishing activity. One-guide faithful NgAgo guide loading... how it’s working in the comments section! Read About NgAgo User's First Impressions Here References ... Additional Resources on the Addgene Blog Read About Cas9 Homolog Cpf1 Check Out High Fidelty Cas9 Mutants...
  17. Deep Dive: qPCR

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    Blog Post
    ...indicating primer dimers. (c) A double peak indicating 2 amplicons. Image credits (respectively): Zuzanna K...details for reproducibility’s sake!   Fig. 2: Setting up triplicates in multiple configurations...used method is the Standard Curve method. See Table 2, (adapted from Wong, 2018) for a comparison of different...Statistically defined log-linear phase   Table 2: A comparison of different qPCR analysis methods. ... for RNA quantitation. Biotechniques. 2000 Aug;29(2):332-7. doi: 10.2144/00292rv02. PMID: 10948434.Taylor... should have a melting temperature 5-10 degrees above the primer and be at least 25 bp away from each ...
  18. How to Negotiate a Successful Lab Start Up Budget

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    Blog Post
    ...identifying the resources needed to accomplish that goal. 2. Identify the tools in that timeline Avoid writing...based on experimental timelines and specific aims. 2. Then make decisions based on financial values. All...Would I be able to get access to these materials?” 2. Always negotiate for a win-win decision An institute...a successful negotiation and collaboration. We’d love to hear more about your experiences and opinions...can get away with using computer modeling and collaborations rather than using animals, then that’s a cost...associated with old equipment, like service repairs, labour costs, and loss of time, can exceed the cost of...Everything is negotiable. Try asking for discounts, collaborations, access to internal award applications, etc...
  19. Antibodies 101: Introduction to Immunofluorescence

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    Blog Post
    ...into the several steps outlined below and in Figure 2. For a deeper dive into the steps and techniques, ...mounting sample on slides, and confocal   Figure 2: High-level overview of an indirect immunofluorescence...recognize the species of your primary antibody and 2) is conjugated to a fluorophore that suits your experiment... Learn about research applications of antibodies in Introduction to Antibodies Learn more about fluorescence...Stay tuned for a future blog post to learn more about these techniques!  Direct vs. indirect immunofluorescence...different approaches to be familiar with when talking about IF, direct and indirect. For direct IF, researchers...immunofluorescence procedure with key considerations about each step. Image created with BioRender.com   The first...
  20. Filming Science Videos in the Age of Social Distancing

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    Blog Post
    ... Figure 2: Ironing boards. Good as a WFH desk, and as a video control center.   Cycle 2: Zooming ...ironing board, giving it the perfect height (Fig. 2). We recorded video using a simple, external video... Production Specialist, I’m constantly thinking about ways to improve our productions to deliver the best...four months after I started with Addgene, I was about to have a check-in with my boss Jennifer. I had ... would be a couple months. Jennifer and I spent about an hour that morning scrambling. Everything we had...someone to be comfortable on camera while thinking about things like lighting, camera angle, and shot composition... allowing people to sit without having to worry about getting in close to mic them up. Monitor: I then...
Showing: 221 - 240 of 618 results