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We narrowed to 675 results for: abo.1

Showing: 281 - 300 of 675 results

  1. Cloning Mammalian Cells with the Agarose Method

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    Blog Post
    ... spread out onto the plate. We test the (capped) 1:1 agarose/DMEM solution against a part of the body ...a 60 mm dish ready for picking is shown in figure 1.   Find Additional Useful Protocols on Addgene's Protocol...specific genes. Many years ago, people in the laboratory became frustrated with using glass cloning rings...the hood. A 48-well plate is also prepared with about 200 ul of culture medium containing serum in each...incubator, additional growth medium is added to about halfway up each well. Tips for using the agarose...with the firmer 2% agarose gel solution discussed above for picking. Many thanks to our guest blogger Iris...

  2. Antibodies 101: Validation

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    Blog Post
    ...expected in another. You’ll need to validate that points 1 and 2 hold true under your specific experimental conditions...application and available resources.     Figure 1: Table summarizing the IWGAV five proposed antibody...like immunohistochemistry on human tissues, when (1) other strategies may not be feasible and (2) large-scale...validation: A view from the mountains. New Biotechnol 45:1–8. https://doi.org/10.1016/j.nbt.2018.08.002  Uhlen...may not be the most accessible option for many laboratories. Additionally, it can be difficult to distinguish...the captured proteins off for mass spec analysis (above). When the results come back it shows that the majority...our Selecting an Antibody blog post! Learn more about Antibodies and their many applications in our Antibodies...

  3. 5 Tips for Troubleshooting Viral Transductions

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    Blog Post
    ...some technical obstacles when working with virus: 1. Viral vectors undergo DNA rearrangements. Depending...approximately 20–50% of the packaging cells after 1–3 days of selection. A lower percentage of surviving...up-to-date tips for your virus experiments. References 1. Anthony, Simon J., et al. "A strategy to estimate...grown at 30 °C instead of 37 °C. If you’re unsure about the integrity of your virus, perform a diagnostic...Additional Resources on the Addgene Blog Learn All about Adenovirus Associated Virus New to Virus? Check ...Check out Our Primer for Beginners! Learn All about Lentiviral Vectors Additional Resources on the Addgene.org...

  4. 22 Hot Plasmid Technologies from 2014

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    Blog Post
    ...expression. Brown et al., Genesis 2014 Jan;52(1):39-48. APEX2 for proteomic mapping and electron...engineering cells that lack release factor one (RF-1; Bacterial strain EcAR7) and minimizing the set of... of human mitogen-activated ERK activating kinase 1 (MEK1) with either one or two phosphoserine residues...comprised of three sets of cloning vectors (Level 0, 1, or 2) which can be utilized in three successive assembly... vectors are directionally assembled into a Level 1 vector creating a single transcriptional unit (Ex:...coding region, and terminator). Next, up to six Level 1 modules can be similarly assembled into a Level 2 ...vectors to allow for additional iterations of Level 1 assembly if necessary. Combining multiple Level 2 ...

  5. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

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    Blog Post
    ...active for greater than 50 generations. Figure 1: Mobile-CRISPRi's modular components. Image from Peters...on a non-selective plate. Most bacteria tested had 1 transfer in every ~100 to 1,000 colonies, but some...including E. faecalis and P. mirabilis, had only ~1 transfer per 1,000,000 colonies. Higher transfer rates...Addgene blog Learn about other CRISPR methods for bacterial genome engineering   Learn about human CRISPRko...

  6. Production of Virus in Insect Versus Mammalian Cells

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    Blog Post
    ...centrifugation (lenti) Centrifugation Titration time ~1 day (AAV); ~1 week (lenti) 2–10 days Initial titers Higher...will help you decide which is right for you. Table 1: Mammalian versus insect cells for viral production... 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 ... can skip the first round of production, saving about a week of work. An important note is that this is...

  7. Designing Your Chalk Talk for the Academic Job Interview

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    Blog Post
    ...acceptable. Aims should be mechanistic, doable by 1-3 people in 1-5 years, and contribute to resolving the overarching...possible, do at least two full length practices (~1 h) and get feedback. It’s also helpful to practice...a single grant. Either way, think realistically about the time and resources required to feasibly accomplish...interrupted frequently by committee members. As mentioned above, plan to give your 10 minute overview and address... list of common questions, check out this eBook about applying for faculty positions. You need to practice...program. You should be proud of it and be excited about getting the opportunity to realize your vision. ...blog posts Resources on Addgene.org Learn more about careers at Addgene Watch our career videos ...

  8. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    ...determined LY6A, also known as stem cell antigen-1 (SCA-1), is the receptor used by PHP.B to reach the CNS...William L. Stanford. "Concise review: stem cell antigen‐1: expression, function, and enigma." Stem cells 25.6... an AAV rookie  Learn about some considerations when using AAVs Learn more about using AAV as a tool for...PubMed PMID: 29175632. If you want to learn more about how PHP.B and PHP.eB were engineered, check out ...Additional resources on the Addgene blog Learn more about PHP.eB, and PHP.S, a peripheral nervous system (...

  9. Tips for arabidopsis transformation

    Type
    Blog Post
    ...with a single insertion because it will segregate 3:1 in the T2 generation. You also have no control over... transgenic plants in the process.     References 1. Emami, S., Yee, M. and Dinneny, J. R. (2013) ‘A robust...transformation’, Plant Methods. BioMed Central, 2(1), p. 19. doi: 10.1186/1746-4811-2-19. PubMed PMID:... 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 ...

  10. 27 Hot Plasmids from 2016

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    Blog Post
    ...a rate of up to 1 per 500-1000 bp. In comparison, the DNA replication error rate is 1 per 109 bp. Unlike...Transcriptional Output-TANGO (PRESTO-TANGO) to screen the NCC-1 library of approved drugs against the entire kit; ...generated 5 common 30bp overlap sequences (Sites #1-5). Each overlap site encodes a unique restriction...First, GMAP-compatible backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed. The authors...is multiplex automated genome engineering (MAGE) (1). MAGE uses recombineering (2) to simultaneously incorporate...of GFP (SEP) by a 9 amino-acid linker (see Figure 1). The key to the biosensor lies in pH: DsRed is relatively...inactivate SEP and leave only DsRed to fluoresce (1). With variants that can be targeted to specific cellular...

  11. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

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    Blog Post
    ...larval stage, Drosophila will pupate, which takes about one week from the initial deposition of the fertilized...there are many resources to help you learn more about fly biology and fly-based experimental systems. ...resources will direct you to the latest information about gene function, while others offer fly stocks and...and other reagents. If you’re ever confused about the best way to study YFG in flies, there is likely an...absolutely essential. You can look up information about your fly gene using the search box in the upper ...In the next post, you’ll learn the fundamentals about how many of these resources work. Many thanks to...Site Specific Recombination in Drosophila Learn about Cre-Lox Resources on Addgene.org Find Drosophila...

  12. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    ...tethered to bring them into close proximity (Figure 1). Formation of the complete FP results in a fluorescent...detected by fluorescence microscopy. Figure 1: How BiFC works. A split FP (e.g., yellow fluorescent... protein interactions in living cells. Nat Protoc 1:1278–1286. https://doi.org/10.1038/nprot.2006.201 ... FP fragments and, preferably, some information about the structures of the tested PIPs. Many split FPs...neurodegeneration. He now spends his time learning and writing about science whenever he's not off the grid.   References...

  13. Better Dyeing Through Chemistry & Small Molecule Fluorophores

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    Blog Post
    ...environments such as live cells and tissues (Figure 1). These flexible strategies give you the best of both...majority of in-cell labeling strategies have two parts: (1) a genetically encoded “tag” expressed as a fusion...Follow him on twitter @rhodamine110.   References 1. Liu, Zhe, Luke D. Lavis, and Eric Betzig. "Imaging... the frontiers of biological imaging.  *Worried about your budget? Email [email protected] Resources on the Addgene Blog Learn About Other Protein Tags Catch Up on Your Fluorescent ...

  14. Tips for Screening with Yeast Two Hybrid Systems

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    Blog Post
    ... fusions that retain their individual activities (1). Researchers also discovered the modular format of...systems could be combined effectively (3). In Figure 1 for instance, the DBD of the fusion protein shown ...ubiquitin system has been devised (8). References 1. Casadaban M.J., Martinez-Arias A., Shapira S.K., ... Plasmids Read our Blog Post About Yeast Vectors Read our Blog Post About Tagging Yeast Genes ...

  15. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

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    Blog Post
    ...experiments or even negatively impact cells (see Figure 1). When behaving properly, FPs enable researchers ... disulfide bonds in the secretory pathway (Figure 1) [4,6–8]. Additionally, FPs that encode N-glycosylation... sarcoma-associated herpesvirus.       References 1. Snapp, Erik. "Design and use of fluorescent fusion...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...

  16. Viral Vectors 101: Parts of the AAV Transfer Plasmid

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    Blog Post
    ...and take a look at each of its parts. Figure 1: Parts of an AAV transfer plasmid. Parts shown in ..., ubiquitous expression, while the human synapsin 1 promoter is neuronal specific (Haery et al., 2019)...the Addgene blog Learn more about basic viral vector elements Read about four considerations for the ...Key elements of an AAV plasmid First let’s talk about the key parts of an AAV plasmid. Inverted terminal...routine plasmid propagation, with complete ITR loss abolishing AAV packaging. To prevent ITR loss, it’s best... Other cis-regulatory elements Next let’s talk about some of the elements that aren’t required parts ...the perfect AAV tool Learn about synthetic promoter AAVs for cell-type expression in retinal cells Resources...

  17. Summer SciComm: Science Storytelling

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    Blog Post
    ...science is all about navigating conflict. In literary terms, conflict can be defined as (1) the struggle...journey even when they know the ending.    Fig. 1: We all know that the superheroes win in the end. ... your science with others.    My favorite quote about science ever comes from an interview in which someone...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 ...

  18. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

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    Blog Post
    ... that dimerizes upon exposure to blue light (Fig. 1). Another blue light-inducible dimerization domain...expression needed for most applications.   Figure 1: With blue light, the VVD domains of the BLADE dimerize...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:... based on the concept of bacteriographs created about a decade ago. To create these images, the scientists...Additional resources on the Addgene blog Learn more about inducible promoters Visit the Plasmids topic overview...

  19. What's the Best Way to Elute and Store Your Plasmid DNA?

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    Blog Post
    .... pH and DNases affect DNA stability Figure 1: So many choices! What to do? The main factors ...your downstream experiment. TE (10 mM Tris-HCl,1 mM EDTA, pH 8.0) buffer is the best buffer for preserving...to purify DNA without a kit Read our blog posts about plasmid cloning Browse our Plasmids 101 blog posts...

  20. Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes

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    Blog Post
    ...optimized for expression in E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2). The key difference... plasmids are available from Addgene! References 1 Bourdeau, R.W., Lee-Gosselin, A., Lakshmanan, A., ...Additional Resources on the Addgene Blog Learn more about in vivo imaging tools The luciferase reporter gene...
Showing: 281 - 300 of 675 results