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Showing: 161 - 200 of 1100 results

  1. New Western Blot Protocol Video!

    Type
    Blog Post
    ...first-timers. That's why we gave our written protocol a video companion! Follow along as Addgenie Meghan and Abi...protocols on our Protocol page, and find our protocol videos on our YouTube page.     ...

  2. An Inside Look into Addgene's Dev Team

    Type
    Blog Post
    ...backgrounds. Sometimes this can lead to wildly different ideas of how to approach a problem, but it is these very...Addgene. Another advantage of Scrum is that it provides transparency to everyone involved about the status...serve Addgene and ourselves. We are also able to consider different technical approaches that may come up...from the user's perspective which in turn helps us decide how to best approach it in our code.  Sometimes...our knowledge, and love to collaborate, which, incidentally, are the same principles for open science. I...

  3. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    ...reinforcement but is considerably simpler than ICSS. The test chamber is divided into two halves that ...s post, we’ll discuss some of the important considerations you should think about when developing optogenetics...the animal perceives through taking a drug. The ideal behaviors to test reinforcement are ICSS and place... of the type of behavior, certain technical considerations still apply. Testing stimulation First, In ... controls without Chr2. c-Fos is a reliable and widely used molecular marker of neuronal activation and...be possible to use c-fos expression to get the evidence you need. For example, as shown in this study ...benefits from collaboration and the exchange of ideas. The rapid spread of optogenetics throughout the...

  4. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    ...endonuclease (Cas9) and a single stranded guide RNA (gRNA). The ~20 nucleotide targeting sequence within the gRNA...reagents are available for genome-wide screens? The goal of a genome-wide screening experiment is to generate...) and "hits" are identified using next-generation sequencing (E). Considerations and tips for successful...underlying genetic cause is not entirely known. Identifying which genes are important for a phenotype can...CRISPR libraries are being used to perform genome-wide screens and highlight some of the reagents that ...to target Cas9 to virtually any genomic locus, provided the target is unique compared to the rest of the...CRISPR technologies, check out Addgene's CRISPR Guide. Figure 1: Like this jar of jellybeans, a...

  5. Inside Addgene's 10 Year Anniversary Gala

    Type
    Blog Post
    ...Science in Boston, where the entire Blue Wing was set aside for the event. As guests arrived, some gravitated... Me Maybe.” After cocktail hour, we feasted on sliders, hot dogs and a mashed potato bar (complete with...well as plenty of delicious desserts. We enjoyed a video highlighting the changes at Addgene over the last... great!) time. Ten years ago, Melina Fan had an idea and her family helped bring it to fruition. The ...Plasmid Sharing Or read more about what happens Inside Addgene   ...

  6. A Quick Guide to a Career in Software Product Management

    Type
    Blog Post
    ...I led. It’s divided into the key takeaway themes and customer pain points we’ve identified. I can’t wait...software product management, I confess that I had no idea what software product management was either. Coming...

  7. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    ...developers of many of these probes, we decided to write this quick guide to help researchers choose the probe...supplied (Baloban et al., 2017).  It is important to consider effective brightness of NIR FPs in cells, in addition...binding of endogenous BV.  When imaging NIR FPs, consider using light sources that produce enough light ...NIR. Usually, mercury lamps are suboptimal, metal halide lamps are better, and xenon lamps are the best ...apparent cellular brightness than previously reported widely used dimeric iRFPs (Table 1). In general, the monomeric...O.S. et al. Single-domain near-infrared proteins provide a scaffold for antigen-dependent fluorescent nanobodies...

  8. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    ...educational science videos is tough. Is the content accurate? Are we conveying complex ideas in the easiest... production standpoint, it wasn’t ideal for our Lab Protocol videos. It’s really nice to have a scientist...perfect height (Fig. 2). We recorded video using a simple, external video recorder and a mic clipped to her...pulled it off! Check out the PCR Primer video! Learn: Producing videos like this would prove to be difficult..., and as a video control center.   Cycle 2: Zooming Forward Design: We produced a video celebrating... cues. We had two options: Film outside at a distance, or film inside with one person in a room. I wouldn...protocol videos?   Figure 4: The remote directing setup, complete with wireless video receiver,...

  9. Single-cell tracking of lineage and identity with CellTag

    Type
    Blog Post
    ...majority of cells failing to develop the desired identity. The development of single cell technologies, ... RNA-seq (scRNA-seq), now allows scientists to identify the gene expression patterns unique to the few...Morris Lab at Washington University in St. Louis provides a combinatorial indexing strategy that simultaneously...simultaneously tracks clonal history and cell identity at the single-cell level, thus linking ancestral gene...Biddy et al., 2018). Tracking lineage and cell identity at the single-cell level with CellTag CellTagging... region. The variable region acts as a unique identifier for individual cells. Each cell is labeled with.... The fixed region acts as a library-specific identifier and allows cells to be tagged with different ...

  10. New Educational Resource: The Addgene Videos Page

    Type
    Blog Post
    ...very excited to debut our brand new Addgene Videos page! Videos are a powerful tool for communicating scientific... first video, featuring Outreach Scientist Jessica Welch, can be found below: How-to videos Our How-To...Videos can save researchers time by quickly relaying key points about an unfamiliar topic or lab process... page, we have neatly organized the educational video content from our website, blog, and YouTube channel...questions you might have about Addgene. The page is divided into three main sections: Protocols To complement...written protocol materials, we have also created video protocols.  These feature step-by-step instructions...various lab processes.  Our most recent protocol video detailing Streaking Plates for Single Bacterial ...

  11. 5 Science Rap Videos & Their Creator: Tom McFadden

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    Blog Post
    ...plasmid rap video for us (If you’re new to plasmids, we highly recommend checking out the video). Tom has...Beautifully filmed video about colony collapse disorder.     5. Plasmid Rap Plasmid rap video made with support...has made many more Science rap videos to teach students around the globe and is pushing SciComm further...Rock Anthem One of Tom's earliest and most popular videos. Describes fossil rocks and their importance in... Check out Our Protocol Pages for New Protocol Videos View Our Molecular Biology Reference Pages ...

  12. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    ...but it isn’t just for lab specimens — there is evidence that lightning can also promote gene transfer ...While these plasmids can be dangerous, they also provide researchers with new means to deliver DNA and other...resistance to toxic substances like heavy metals or pesticides, enabling their hosts to survive where others... T. M., & Simonet, P. (2001). Laboratory-scale evidence for lightning-mediated gene transfer in soil. ...

  13. Identifying Sequence Elements with SnapGene's Feature Database

    Type
    Blog Post
    ...database of common features, and devising rules for identifying a feature even when the match is imperfect. SnapGene's...catalog every variant of a feature. Instead, we identified common variants, and then crafted a detection...reasonable rule is to require at least 96% sequence identity when detecting a reference feature. For a coding...match even when the DNA sequence has changed. Identifying plasmid control elements Coding sequence features...original literature, some of it decades old, to provide reliable feature annotations. Even with these extensive...that have different properties. To prevent misidentification, we augment the database with new fluorescent...protein variants. The future of DNA feature identification What’s next for common features? We plan to...

  14. Transferable Skills Guide: Career Planning Resources

    Type
    Blog Post
    ... it! Further Reading and Outside Resources http://www.businessinsider.com/how-to-get-the-most-out-of-job-fairs...your lab, right? Wrong! While it is clearly a good idea to be diligent in your research, remember that you...themselves, resources are often scarce and difficult to identify. So what specifically should you do? What follows... elaborated on later. As for what job fairs to consider, some of the best options are the Naturejobs Career...most relevant experience at the top. You may also consider visiting any of the employer’s career pages to... apply for the position. This post on Business Insider nicely summarizes good job fair practice. Career...and other online career resources Once you’ve identified places you’d like to work, get online and take...

  15. A Guide to Starting Your Own Journal Club

    Type
    Blog Post
    ...journal club. Sorting out the logistics Once you decide on the topic, there are a number of administrative...notice for the first meeting so that people can set aside time in their schedules and read the paper (PS: ... on single papers could be more important.  Any slides, presentations, recordings, and other materials... ensure that the journal club is useful for the widest audience, you should get feedback from those who...everyone to offer thoughts you may not have even considered. These questions should be tailored to the group...priority for the participants. Perhaps people feel confident in their own knowledge of the field. Perhaps interest... not actively participating, it may be time to consider ending the meeting. This is a common failing in...

  16. Transferable Skills Guide: Cross-team Communication

    Type
    Blog Post
    ...2) creating a pictorial guide for our engineers (Fig. 2). Creating this guide was relatively easy. I took...different interpretations of what we discussed and decided. I needed to a) figure out the sources of miscommunication...non-scientist friends and family is going to vary widely. The way you communicate your science at a poster...collaborations with other scientists within and outside your field, and any number of interactions with...the context you do, which means you’ll need to provide that context yourself. Simple words are not so...the same question and asking can help the team identify possible points of confusion in the future. Improving...

  17. A Guide to Designing a Scientific Poster: Content, Layout, and More

    Type
    Blog Post
    ...questions, data, and conclusions should be clearly identified and interpreted in less than a couple of minutes...Tips for designing your poster Check conference guidelines To begin, you should always follow conference...poster at all times, so other participants can identify you. You may also wish to make creative use of...three, or four column style, and in all cases the idea is the same; the reader will navigate the poster... to low, some of the poster will naturally sit outside of the reader’s field of view because of its length...to read. The Thompson Rivers University Library provides useful tips on how to craft this type of poster...doctoral student at Michigan State University. The idea is that the main research finding is written across...

  18. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    ...reverse transcriptase) together with a prime editing guide RNA (pegRNA). The pegRNA consists of a target-complementary...on the website or offline via GitHub. The model provides users with different designs of pegRNAs, ranked...and test them in their labs. Figure 2: Guidelines to predict pegRNA efficiencies with pridict.it...computational prediction models. In addition, we provide options for nicking gRNAs (used for PE3 and PE5...self-targeting screens such as PRIDICT/PRIDICT2.0 provide valuable insights into pegRNA editing efficiencies...Brinkman, E. K., Leemans, C., Vergara, X., van der Weide, R. H., Morris, B., van Schaik, T., Manzo, S. G....Addgene.org CRISPR: PRIME Edit Collection The CRISPR Guide Empty gRNA Expression Vectors...

  19. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ...protein of interest, these tools provide an easier means to identify targets through fluorescent labeling...Lavis, L. D. (2022). Caveat fluorophore: an insiders’ guide to small-molecule fluorescent labels. In Nature...interiors to many inquisitive scientists. Seeing inside these specimens has offered illumination on biochemical...drug development (Grimm & Lavis, 2022). With the wide breadth of fluorescent tools that have been developed...will outline some fundamental characteristics to consider when selecting a dye or ligand to make your selection...protein, enabling the labeling of protein substrates/peptides and antibodies. Example applications of this include...is bound through green light emission. Key considerations in selecting fluorescent dyes/ligands Effects...

  20. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    .... The video abstracts provide a brief introduction to the research and, after watching the video to determine...platform that provides links to the videos, keeping the copyright and control over the videos with the researcher...others. Improving science communication through video     Video abstracts about research, especially published...view count and videos published, YouTube reported a total of 865,995 views for the 926 video abstracts identified...attention for their work, a video abstract may be a useful tool for turning video views into online article...few years, I decided to look at what online tools people generally like to use to share ideas – with the... online video market ever since. The take home message of all of this appears to be that video is the ...

  21. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    ... CRISPR RNA Scaffolds." Adam has also created a video to help scientists visualize the concepts discussed...endonuclease activity coupled with a customizable single guide RNA (sgRNA) that bears complementarity to a 20bp...RNA (scRNA) will bind to a PAM-adjacent target nucleotide sequence then subsequently recruit a transcriptional...orthogonal dCas9 protein that will still recognize guide RNA but with a different dCas9 binding module, alternative... molecular microbiologist. Summary  The single guide RNA in the CRISPRi system can be repurposed to include... Qi, L. S. et al. Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene ...

  22. Quantifying DNA? Here are Five DNA Quantification Methods to Consider

    Type
    Blog Post
    ...things to consider: cost, time, equipment, and expected DNA concentration. It’s important to consider the advantages...DNA sample.  These absorbance measures give you an idea of the concentration of your DNA prep and whether...A280 ratio is used as an indicator of DNA purity. Ideally, this number should be between 1.8 and 2.0. The...containing a DNA intercalating dye (ex: ethidium bromide) and choosing a DNA ladder with known concentrations...

  23. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...FLEx Vectors Important Considerations When Using AAVs Three Key Considerations For Precise Neuronal Targeting...reproducibility of the data and can instill greater confidence when interpreting the outcome.    References...Sequencing and Analysis Platform to Assess the Identity of Recombinant Adeno-Associated Viral Preparations...

  24. Quick Guide to Working with Drosophila Part 3: Genome Engineering in Flies

    Type
    Blog Post
    ...gene function. In this final installment, I’ll provide a brief introduction as to how you can engineer...Addgene also has a number of insect vectors to consider. An easy way to see what is available is to browse...transgenic fly There are a few important points to consider when choosing the vector you’d like to use to ...pathogens and their hosts. It’s not always clear which side he roots for. You can try to convince him to use...

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

    Type
    Blog Post
    ...deposition of the fertilized egg by its mother. Inside the pupal case, the larva will metamorphose into...light in coloration and can look very plump. On the side of their abdomen will be a dark spot. This is meconium...visible through their translucent bodies. This is the ideal stage to select for females because you can be certain...have an allele that causes the hairs on the dorsal side of its thorax to look like stubble (sb). For a better...search box in the upper right hand corner. They have video tutorials under the help tab like this one to get...pathogens and their hosts. It’s not always clear which side he roots for. You can try to convince him to use...

  26. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...have allowed scientists to easily perform genome-wide screens to effectively and efficiently investigate...libraries contain many sgRNAs per gene to increase confidence in the genetic screen. However, there are benefits...utilize only a few sgRNAs per gene. These libraries provide more information while using less resources and... the Broad Institute have developed human genome-wide CRISPRko (knock out), CRISPRi (interference), and... sgRNAs that were designed based on the rules identified in Doench et al., 2014 to predict sgRNA activity...scientists examined approximately 1,900 sgRNAs and identified sequence features and adjustments that improved...performance. The Root and Doench lab’s optimized genome-wide CRISPRko, CRISPRi, and CRISPRa libraries have been...

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

    Type
    Blog Post
    ...the Dark Side." --Darth Vader in "Return of the Jedi" While Vader was referring to the evil side of a mystical...and rewarding. But, what if an FP fusion with a resident endoplasmic reticulum (ER) protein instead localizes...Previously, we have described a number of practical considerations that should be taken into account when deciding...must therefore determine your cloning strategy and decide whether you will create an N- or C-terminal fusion...needs. Equally important, before cloning you must consider whether and how well your FP will function in ...secreted into the extracellular milieu. Many of the resident proteins within these compartments undergo significant...post-translational modifications including glycosylation, disulfide bond formation, and proteolytic cleavage. FPs ...

  28. CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

    Type
    Blog Post
    .... Watch the following video to learn Daniel and Matthew's procedures. The video details how to transfect...Harvard Medical School. Addgene is proud to present a video reprint of the CRISPR article "Generation of Genomic...the Journal of Visualized Experiments (JOVE). The video publication by Stuart Orkin and Daniel Bauer's labs.... Biallelically disrupted cells can be easily identified. Even in the case of clones with monoallelic ...for biallelic deletion clones:    Like the video? The Orkin and Bauer lab's related paper describes... as their techniques for creating deletions, identifying these deletions, and finally characterization...

  29. SpyLigase Irreversibly Locks Peptides Together for Efficient Cell Capture

    Type
    Blog Post
    ...just join one peptide tag to another peptide tag. Addgene: What is the advantage of peptide tags for building... locking two peptide tags together. SpyLigase overcomes limitations in the use of peptide tags, which ...assemblies? Howarth: Peptide tags can be very generic; you should be able to use the same peptide tag for all ...thanks for the T-shirt! References: SpyLigase peptide-peptide ligation polymerizes affibodies to enhance ...itself together, to produce irreversible protein-peptide interactions. In a study published in Proceedings...Developing a new tool Addgene: Where did you get the idea to develop SpyLigase?  Howarth: It came from our...the Streptococcus pyogenes protein domain made it ideal for this purpose? What led you to it? Howarth: Our...

  30. From our Table to Yours: An Inside Look at Lunch at Addgene

    Type
    Blog Post
    ...four of us would eat together every day, sharing ideas for the company, but also talking about sports, ...Addgene. We’re a small non-profit, so we can only provide free food on special occasions. However, employees...and make time for special projects? We borrowed an idea from the software folks and started Hack Days. During...plasmid protocols for our website. Free lunch is provided for participants during the Hack Days, and we’...Lunches” With more employees, Addgene has naturally divided into smaller functional teams. Our Executive Director...make time for free food, and these lunches would provide a friendly setting for people in different labs...Who knows what could come out of it? Perhaps new ideas or collaborations, or at least a shared sense that...

  31. HA Frankenbody, a New Imaging Tool to Visualize Single Molecules and Nascent Peptides

    Type
    Blog Post
    ...living systems, and often do not fold properly once inside cells.  Developing the HA frankenbody To combat...genetically encoded antibody based probe that detects the widely used HA epitope in diverse cellular environments...epitopes (Ewert et al., 2004). This scFv hybrid provides the probe structure and stability from the scaffold...translated quickly by the ribosome. This feature provides HA frankenbody the ability to bind nascent HA ...a versatile tool that can be used to visualize a wide range of protein dynamics in vivo. In the future...Resources on Addgene.org Read our fluorescent protein guide Find plasmids for subcellular localization Find ...

  32. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

    Type
    Blog Post
    ...practices and guidelines. It was wonderful to see the development of this practical guide by the workshop...networks, and specific neuronal populations to consider. To tease apart how different neurons impact a... discussions as a publicly accessible practical guide which was recently published in Frontiers in Neuroanatomy...experimental approach you choose, it’s essential to consider the technical limitations of each approach and...science.   Check out the precise neuronal targeting guide here! Additional resources on the Addgene blog ...retrograde AAVs Resources on Addgene.org Read our AAV guide Learn more about our viral service Find retrograde...

  33. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

    Type
    Blog Post
    ... organization, contact me! I can provide a step-by-step, how-to guide to get one started. Advantages ...communication skills. The leader also provides materials such as pre-reads, videos or exercises to be completed...increases the diversity of input and perspectives Provides a good mechanism for accountability that makes...to talk about?” There are an infinite number of ideas on the internet, but start by downloading my handy...collected. The next blog in this series will have more ideas on what to do once you are in a mentoring relationship...Peer Mentoring relationships? Comment below with ideas from your own mentoring group or with any questions...

  34. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

    Type
    Blog Post
    ..., Moullier P, Ayuso E, Vandenberghe LH (2017) AAV-ID: A Rapid and Robust Assay for Batch-to-Batch Consistency...sequencing (NGS) QC platform that allows us to provide scientists with complete plasmid sequences. After... viral genome sequencing (VGS), to confirm the identity and serotype of our AAV preparations. We recently...an effective tool to confirm AAV vector genome identity. These studies used SMRT sequencing, TA-based ... that VGS can quickly and reliably confirm the identity of AAV genomes and detect contaminating AAV DNA...packaging cell line, helper and packaging plasmids. We decided to use this to our advantage by designing a Python...from multiple AAV lots can be run in parallel and provide serotype data in minutes and the scripts  can be...

  35. Download Addgene’s Science Career Guide, Plus 8 Career Tips from Addgenies

    Type
    Blog Post
    ...one comprehensive, downloadable guide: the Addgene Science Career Guide. We also asked Addgenies what their...endeavor.  Break the process down into steps and set aside time during the week to tackle each one.  Get the...you’d like to see discussed on the Addgene blog, consider writing a guest post. Additional resources on...careers blog posts Read our transferable skills guide Resources on Addgene.org Find job listings at ...

  36. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    ...safe-targeting guides vs. non-targeting guides on screen analysis and hit calling. 5,644 non-targeting guides and...short guides had reduced toxicity compared to full-length guides. Reduced toxicity of short guides could...genes identified with short and long guides. Together these results suggests  that short guides have fewer...demonstrate safe-targeting guides as a potential control, and provide evidence that truncated gRNAs have...Genome-wide CRISPR/Cas9 screens are a high-throughput systematic approach for identifying genes involved...biological process. These screens provide an alternative to genome-wide RNAi screens, which although highly...questions about genome-wide CRISPR/Cas9 screen design Are non-targeting guides an appropriate control...

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

    Type
    Blog Post
    ...choice of junction sequences guided by experimentally-derived DNA Ligase fidelity data. Our work demonstrates...accuracy of assembly. The fidelity data can be applied to derive similar high-fidelity overhangs for any Golden...Reactions are often cycled between 37°C and 16°C to provide optimal reaction temps for both the restriction...appropriate levels of colony plating densities. Ligase fidelity and assembly efficiency Five fragment lacI/lacZ...redesigned for 12 and 24 fragments.  This redesign was guided by both advances in the re-engineering of the original...restriction enzyme and the completion of T4 DNA Ligase fidelity studies conducted by Potapov et al. at NEB (1,...Recently, NEB researchers profiled the comprehensive fidelity of cohesive end ligation by this enzyme for all...
Showing: 161 - 200 of 1100 results