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Showing: 241 - 270 of 954 results

  1. Bringing Sustainable Practices to the Lab: Innovations

    Type
    Blog Post
    ...New ideas and approaches are encouraged in science. It isn’t surprising that new ideas are also popping...lab lives. Read on to learn about some creative approaches to lab sustainability, and find ways to further...but what about all the manufacturing steps that happen before you even receive the product? How can we...tips. This is part of a cultural change in how we approach waste that needs to be supported at all levels... facility, where it’s sanitized through an EPA-approved process and no longer considered hazardous. Then...industrial and residential markets. Rethinking our approach to lab waste is something we all need to do. Remember...

  2. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ...Early CRISPR applications were often limited by the low editing efficiency of homology-directed repair...BE3, nicks the unmodified DNA strand so that it appears “newly synthesized” to the cell. Thus, the cell...reduce the undesired C→G or C→A conversions that can happen with BE3s. These byproducts likely resulted from...making them a promising tool for cell therapy applications. Dual base editors What about combining the ...Grünewald et al., 2020). Another lab took a similar approach. Dali Li’s lab’s A&C-BEmax consists of a fusion...activity compared to ABEmax. In an alternative approach, engineering and directed evolution of the TadA...editors with increased activity and therapeutic application. Nature Biotechnology, 38(7), 892–900. https:...

  3. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    ...of the protein of interest. An electric current applied to the gel causes the negatively charged proteins...with blotting paper and an electric current is applied causing proteins to migrate out of the gel and ...sandwiches.   Figure 2: The western blot dry apparatus includes a stack of filter paper, a copper anode...search, first pay attention to the validated applications listed on the vendor’s website and choose an...western and vice versa.  In addition to validated applications, pay close attention to the controls listed ...choose antibodies that are validated for your application and test the antibody in your specific system...to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences...

  4. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ...attB-attP pairs from different recombinases are overlapping or nested on a register, then the operation of...more population-wide (as opposed to single-cell) approach is to interrogate state with qPCR. The re-arrangement...performed on one register at a time. If a particular application makes it difficult to isolate single registers...state read-out method may not be suitable for applications that require minimal cellular perturbation. ...contains the register. Depending on the desired application, the inducible promoters we used to drive recombinase...program to find the one that best suits your application. How efficiently do RSMs work? We tested a two-input...anticipate that, depending on the context and application, some inducible recombinase systems may work ...

  5. Year of the Camelid: Antibody Style

    Type
    Blog Post
    ...diagnostic applications. For example, fluorescently and radioactively labeled sdAbs have become appealing candidates...to sdAbs becoming valuable tools for diverse applications. For example, RANbodies can be used as an alternative... as well. The first sdAb based therapeutic was approved in 2018, with many others in clinical trials (...maintaining a llama facility is out of reach and unappealing to many labs), the field is moving towards using...inspired this versatile family of tools, the applications they have already enabled, and the discoveries...

  6. We're Updating Our Hold for Publication Policy

    Type
    Blog Post
    ... complete necessary quality control and legal approvals so the plasmids are ready for distribution as ...this Hold for Publication policy seriously and appreciate the trust you’ve put in us when you choose this...worry, we’ll let you know before these releases happen (and congratulate you on your paper!). And as always...your deposit or plasmid ID numbers and we’ll be happy to update the plasmids to add the citation and release...hope somebody will find the plasmids useful, we’re happy to distribute unpublished materials. What about ...needs, so please let us know what you think. We appreciate your feedback. If you have questions about depositing...

  7. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    ... be used as a fusion tag for a wide range of applications from traditional imaging to FRET and stochastic...transforming E. coli with a gene library of approximately 484 variants, including some potential tetramer-breaking...Florida State University, they used a mutagenesis approach and found that specific amino acid changes favored...violet range. For example, mWasabi can be used with Sapphire (excitation maximum 399 nm) because it does not...fluorescent protein: structural characterization and applications in fluorescence imaging." Biochemical Journal...identification of the molecular determinants of color and applications in fluorescence imaging." BMC biology 6.1 (2008...

  8. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    ... Many thanks to our guest blogger, Erik Snapp! Erik Lee Snapp received his PhD in Molecular Microbiology...This post was contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at theJanelia... have been completed. Therefore, to be ready to apply for a Group Leader position, key first author manuscripts... project. Furthermore, Group Leader position applications require 3 letters of reference- the graduate...informal recommendations, more positive grant application and manuscript reviews, etc. You'll also meet... fluorescent proteins,  and live cell imaging approaches. Erik is also an avid long distance runner, gardener...

  9. Important Considerations When Using AAVs

    Type
    Blog Post
    ...possible tools, lentiviruses for example, are not approved for use in vivo use at the University of Manitoba... be mostly on the brain, but many of these tips apply to all tissues. Subscribe to Viral Vector Blog Posts... infect) can influence an AAV’s value greatly. Applying this mantra to AAV selection, there are three ...of AAVs? 1) Combined functional and anatomical mapping of circuits In our lab, we use AAV-DREADD complexes...only can we compare these methods in specific applications, we may also be able to use these comparisons... on overall cell function. AAVs are the most appropriate vehicle for this type of research, as they are...efficiency, and safety profiles should strengthen its appeal for the scientific and medical communities. Many...

  10. Predicting Adverse Reactions to Monoclonal Antibody Drugs

    Type
    Blog Post
    ...   Limitations of the in silico approach The in silico approach cannot replicate the complexity of ...Better together Combining in silico and in vitro approaches enables researchers to develop safer and more...unwanted immunogenicity. The 3-D models can be applied to corroborate the occurrence and extent of the... somewhat act like a 2-step verification. All approaches have their own limitations. Using methods together...together can inform decisions about in vivo application and clinical testing. This would pave the way for...

  11. "Build Your Plasmid": The Game - Play to Teach

    Type
    Blog Post
    ...information they’ve absorbed if they don’t get a chance apply it in an interactive setting. The problem is…how... as I try being compelling, interactive, and application-driven to boost curiosity, I have never experienced...they would use their logic and strategy skills to apply those concepts and understand how therapeutic strategies... BUILD YOUR PLASMID is a game where you need to apply basic molecular biology, cell biology, microbiology...best strategy and propose a final gene therapy approach for the disease to be openly discussed with the...opposing team. In the end, the students had fun while applying most of the concepts required by the course and...

  12. Science Career Options

    Type
    Blog Post
    ...critically and carry out in-depth research is applicable to a wide range of industries and is a highly...skills acquired from a science degree that can be applied to many roles. These types of skills show that ...sectors such as forensics currently have too many applicants compared to the number of available roles). Make... Make your resume stand out Before you begin to apply for a job, it is useful to search online job websites...your research, it’s time to start sending out applications. Check out our other career posts for more specifics...careers and to find advice on crafting the perfect application. Additional Resources on the Addgene Blog Learn...

  13. Management for Scientists: Delegating is Key

    Type
    Blog Post
    ...dramatically improved when tasks are distributed appropriately. If team members have opportunities to develop...develop and stretch then they will be happier and more productive in the long run.  If you are overwhelmed...the copy, design every icon, curate all of the appropriate plasmids, and code the entire page. That would... page. The team manager must then facilitate appropriate communication between members of the team to ...some important criteria for delegating tasks appropriately: Are the desired goals and outcomes clear? ...questions. If you are perceived as too busy to be approachable mistakes will be made that could have been prevented...

  14. Viral Vectors 101: Types of viruses

    Type
    Blog Post
    ...cell type doesn’t always lend itself to direct application of virus. However, the cargo capacity of AAV ...gamma-retro viruses. This is large enough for many applications, but in some cases multiple AAVs (co-infection...and AAV. If you’re wondering more about what applications these viruses are good for, we have plenty of...Adenovirus-Mediated Gene Delivery: Potential Applications for Gene and Cell-Based Therapies in the New...Gammaretroviral vectors: biology, technology and application. Viruses. 2011 Jun;3(6):677-713. DOI: 10.3390... Campos MD, Materatski P. An Overview of the Application of Viruses to Biotechnology. Viruses. 2021 Oct...

  15. AAV Q&A with Tim Miles

    Type
    Blog Post
    ... our knowledge. Disease Applications Q13: Does AAV have potential applications in diseases which may be... be best served by a dual vector, split intein approach.   Q3: What’s the best method for titering AAV...1x1014 vg/ml.. As evident in our repository, this application is an outlier for PHP.eB (1x1011 - 1x1012 vg/...systemic injections? A: We have not observed any appreciable difference in AAV transduction efficiency between...CRISPR tools is an exciting area for future AAV application to epigenetic editing. This area is discussed...

  16. Hot Plasmids - February 2022

    Type
    Blog Post
    ...editing applications.  Figure 4: Overview of CasMINI for genome engineering applications. Image ...stability in cells, and use it in a variety of applications. This set of new anti-FLAG frankebodies includes...to FLAG-tag H2B for nuclear localization. (c) Applications of anti-FLAG-FB-GFP labeling various targets... and anti-HA frankenbodies have a variety of applications, including live-cell imaging, single-molecule...challenges for in vivo delivery and cell engineering applications. CasMINI, a new miniature CRISPR-Cas system ...

  17. How to Keep a Lab Notebook for Bioinformatic Analyses

    Type
    Blog Post
    ...information in the future. Approach: Outline a brief overview of the approach that you will take to help...biologists are finding themselves using computational approaches to analyze large data sets (myself included) ...your analysis, even if your conclusion is “this approach is not ideal because…” Including a conclusion ...or may not be where you also record your goal, approach, and conclusions (see above). Keep original ...in my own research. Again, some of this may not apply or may not work for you. The best way to find a ...

  18. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    ...fluorophores, together, are able to give you the appropriate readout. A common method of labeling proteins... wavelength. Likewise, the emitted photons also appear within a range of wavelengths. For that reason,...excitation and emission are the most efficient (i.e., happen most of the time). Figure 1 shows a graph from ...higher-wavelength emission spectrum compared to APC. This happens due to a process called Fluorescence Resonance ... the fluorophore, and whether that fluorophore appears alone or in conjugation with another fluorophore...absorption at 650 nm, APC-Cy7's optimal excitation happens at higher wavelengths of about 750 nm. Source: ...Besides some background cell death constantly happening in cell culture, the transduction process itself...

  19. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    ...upcoming posts in this optogenetics primer series, I’m happy to share some of the tips and tricks and I’ve learned...memory task, on the other hand, would not be an appropriate test.  Of course, proper controls are required...using these lines is, if one is crossed with an appropriate mouse line expressing a particular cre, viral...22430017. 2. Fenno L, et al. The development and application of optogenetics. Annual review of neuroscience...PMC3225502. 4. Mattis J, et al. Principles for applying optogenetic tools derived from direct comparative...neuronal populations using combinatorial viral approaches. Current protocols in neuroscience / editorial...PMC3191888. 9. Zeng H, Madisen L. Mouse transgenic approaches in optogenetics. Progress in brain research. ...

  20. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

    Type
    Blog Post
    ...developed by the James and Schuh labs. Clift et al. applied TRIM-away to nine proteins in ten ddifferent cell... sometimes incorporating chemically inducible approaches to provide temporal control, this new system,...Epigenome editing by Cas9 (FIRE–Cas9), combines these approaches to enable rapid and reversible recruitment of...target multiple loci at once. In addition, the approach allows minute-by-minute analysis of the kinetics...domain. Transcription factor activation via the appropriate ligand - estradiol for hER or progesterone for...leads to nuclear localization, binding to the appropriate promoter sequence and activation of gene expression...

  21. Meet an Addgenie: Insights into Grad School, Career Paths, and More

    Type
    Blog Post
    ...the co-op application process, my advisor encouraged me to keep my values in mind as I applied. Thinking...My name is Lauren Flynn and I am currently wrapping up my co-op with Addgene as a DNA Sequencing Analyst...s and leave. I was really devastated when that happened. I didn't know if I should even stay in science...in your career path and still find comfort and happiness there. Read all of the Meet an Addgenies features...connections with my coworkers that may not have happened if we hadn’t had these conversations. Now, I am...

  22. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    ...simpler, yet more effective, CRISPR-based epigenetic approach (spoiler alert: it’s dCas9… with no modifications...). But there are a number of drawbacks to this approach: TET proteins are not demethylases; they are ...and Szyf 2021).    Figure 3: A simpler approach - dCas9 blocks DNMT1 methylation, leaving the ...resistance genes.  It’s that simple! Good luck and happy editing!    Thank you to our guest blogger!Daniel...Addgene blog CRISPR101: Editing the Epigenome  Mapping the 4D Nucleome with CRISPR/Cas9 Resources at ...Xu X, Tao Y, Gao X et al. (2016) A CRISPR-based approach for targeted DNA demethylation. Cell Discov 2,...

  23. Synthetic promoter AAVs for cell-type specific expression in retinal cells

    Type
    Blog Post
    ... Ophthalmology Basel recently took a different approach to enhance specificity in retinal cells. To do...type specific expression in the retina.  Four approaches for designing synthetic promoters  To build a...reproducibly labeled retinal cells can be found here.  Applications for synthetic promoter AAVs Besides restricting...synthetic promoters have a few other potential applications: Using promoter combinations for targeted expression... promoters expressed in. The lab tested this application using one promoter to drive expression of Cre-mCherry... rates of labeling are ideal of gene therapy applications, promoters that produce sparse labeling would...

  24. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ... to increase AAV transduction efficiency by approximately 15-fold (Chen et al., 2006). 3. Transduction... at 4 °C). After ultracentrifugation, virus may appear as a white pellet. The pellet can then be rinsed... This antibiotic selection can be performed approximately 72 hours after transfection and should give ...give rise to a surviving population of approximately 20–50% of the packaging cells after 1–3 days of selection... could be examined with a microscope and should appear fluorescent if they were infected. While there ...transfection, higher-level expression after approximately 72 hours is likely the result of successful ...

  25. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...to be less expensive than an ELISA kit. In this approach, viral RNA is first converted to cDNA and then...several different lentiviruses in parallel, this approach should be considered. Similar to the p24 assay... Fluoresence Titering Protocol Another popular approach utilizes the antibiotic resistance genes carried...have integrated into a target cell’s genome. This approach can discriminate multiple integration events, ...underestimating the viral titer. The main drawback of this approach is that it can be quite labor intensive. Target...virus may want to choose more straightforward approaches such as the FACS-based or colony-forming unit...

  26. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    ...18% to 65% (Maggio et al., 2014). An alternative approach to NHEJ is to utilize the Homology Directed Repair...phenomenon to the presence of terminal proteins capping the ends of linear double-stranded AdV genomes....double-stranded chromosomal DNA breaks “that always happen naturally.” These adenoviral CRISPR/Cas9 genome...and quiescent, post-mitotic mammalian cells. Applications of CRISPR delivery by adenoviral vectors Since... Adv vector delivery is often used, the CRISPR approach based on HDR has the advantage of being a potentially...Strategies for Tissue-Specific Targeting. In: Applications of viruses for cancer therapy. Elsevier, pp ...

  27. Of Myc and Men

    Type
    Blog Post
    ...The strategy they landed on was to recombinantly append their proteins of interest with unrelated peptides...generate a reliable antibody for protein tagging approaches” was not part of the goal when Evan’s team generated...plays in human health and disease. But no science happens in a vacuum!  Munro and Pelham learned of these... flexible - it could now be used in way more applications.   Fig. 1 Making a Protein Tag (Abridged...their study, some of the newer Myc-tag antibodies appeared to be less sensitive to sequence context.    So...proteins expressed from cloned genes: deletion mapping functional domains of Drosophila hsp 70. Embo J...

  28. Academic vs. Industry Postdocs

    Type
    Blog Post
    ...your resume pulled to the top of the pile when you apply for such positions. Of course, if your goal is to...transition from an academic setting—especially if job applications require teaching experience or high numbers ...web site. Each position is for a specific, pre-approved project. Other companies, such as Novartis, have...s program. Once you’ve found a few positions to apply for, compare and contrast industry postdocs at those...not. The point is that each company has a unique approach, and some might suit your needs better than others...background in cell and developmental biology and is applying this towards researching neurodegenerative diseases...

  29. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    ...10 modules often rely on two-step hierarchical approaches using different Type IIS restriction enzyme specificities...transformation plate can be manipulated to result in appropriate levels of colony plating densities. Ligase fidelity...accuracy of assembly. The fidelity data can be applied to derive similar high-fidelity overhangs for any...researchers continue to test the limits of DNA assembly approaches with increasingly complex experimental conditions...Kucera is a Principal Development Scientist of Applications & Product Development focusing on DNA cloning...Optimization of Golden Gate assembly through application of ligation sequence-dependent fidelity and bias...

  30. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

    Type
    Blog Post
    ...and Mar. 1, 2020. Figure 2: A simple approximation of the effect of COVID-19 on data smoothed with...a 7-day rolling mean. While this global approximation already shows us a rough "COVID-19 effect" on...maximum likelihood estimates of each parameter. We approximated the data likelihood with multivariate Gauss-...the supposed end of lockdown), request activity appears to return to normal. The model fit these two discrete...is a Ph.D. candidate at MIT. His research uses applied machine learning and statistical methods for fundamental...fundamental scientific discovery and high-impact applications.       Alvin Hsu is a graduate student at Harvard...
Showing: 241 - 270 of 954 results