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Showing: 481 - 520 of 1023 results

  1. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    ...digital notebook and make sure you note the freezer location of every plasmid in your collection. Use one of...access and use the materials they need, and avoid duplicating your effort. In addition, if you should choose... important step in ensuring quality, as it will catch any errors made during the cloning, sample handling... for more tips on sequence analysis, plasmid verification, and using NCBI BLAST. Tips for high-throughput...result against the expected sequence for quick verification, which can be done through the align tab of ... plasmid verification...

  2. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog Post
    ...As evidenced by all the CRISPR publications, press, and plasmids out there, it’s obvious that CRISPR ...nickase variant, lowering Cas9 expression, and truncating gRNA sequences used for targeting; however, these...on-target and off-target activity at multiple genomic locations in HEK cells. Further testing of two of the mutants...PubMed PMID: 26628643. Find plasmids from this publication at Addgene. 2. Nishimasu, Hiroshi, et al. "Crystal...PubMed PMID: 26735016. Find plasmids from this publication at Addgene. 5. Chen, JS, et al. "Enhanced proofreading...PubMed PMID: 28931002. Find plasmids from this publication at Addgene. Resources on the Addgene Blog Read...

  3. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    ...repurpose popular GFP reporter lines for more sophisticated experimental manipulations, saving the time ..., 98% of TdTomato expressing cells were GFP+, indicating a robust, but specific system. T-DDOGs also successfully...systems are easily adaptable to neuroscience applications, including optogenetics, and they should make...Could Cas9-DOG be developed soon to allow genome modification only in a small subset of cells? Other proteins... PMCID: PMC4096992. Find plasmids from this publication at Addgene. 2. Tang, Jonathan C.Y., et al. “...PubMed PMID: 26258682. Find plasmids from this publication at Addgene. 3. Kirchhofer, A., et al. “Modulation...

  4. Career Coaching for Scientists: Why and Where Do I Find One?

    Type
    Blog Post
    ...choose well. What are their qualifications? Do they have certifications? How can they demonstrate their...  Check out Joanne's Reddit AMA I am a strong advocate for peer mentoring, but volunteer mentors are ...help you. A good coach usually will be a clear communicator and make direct suggestions for improvement ...coaching is nice if you can get it.  Because I am located in Cambridge, MA (and because we have a lot of ...she is especially excellent in coaching for communication skills. Industry transition is also an area ...

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

    Type
    Blog Post
    ...scientific poster apart? First, a good poster should catch the audience’s eye and draw people in. That can ... sections should be as concise as possible, communicating only the most important results to your audience...have the advantage of being able to verbally communicate your work, so you don’t need every little detail...may choose to display it as rows. It can be more catchy because it is generally less common and will stand...advantage of being different and potentially more catchy, but it can be trickier to make figures fit nicely...relevant to the work you are presenting. The latter is catchy, but beware. A distracting background or clashing...

  6. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ...experiences to undergraduate students (1, 2), but educators face significant challenges in designing suitable... become very different due to a whole genome duplication and extensive rearrangements in the S. cerevisiae...overexpression vector. This plasmid contains origins of replication and selectable markers that allow students to...met deficiencies (manuscript in preparation), indicating that Met protein functions have been largely ... research into undergraduate courses and in communicating science to the public. You can find out more...). Vision and Change in Undergraduate Biology Education: A Call to Action, Washington, DC. 2. President's...

  7. Hot Plasmids February 2024

    Type
    Blog Post
    ..., its dimeric nature can interfere with some applications. Several labs have been working to produce monomeric... for molecular fusion and membrane-targeting applications. Nature Methods, 1–9. https://doi.org/10.1038...library generation and screening that led to identification and characterization of AAV.CAP-Mac. Image ...rodents and broad tropism in primates. Nature Communications, 14(1), 3345. https://doi.org/10.1038/s41467...proven itself a good option for essentially all applications. Addgene’s plasmid collection includes over ...clone (M2) has been widely used in a variety of applications. Find anti-DYKDDDDK recombinant antibodies here...

  8. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    ...the best, or even which one will work for your application? Like many scientists, I have spent hours sifting...produced from a new set of animals), and wrong application (different experiments/conditions can change ...differently over time), compatible with the desired applications (such as immunohistochemistry (IHC) or western...colleagues! Dr. Wu and his lab members (like me!) are dedicated to creating a public and accessible database for...expectation and comparison on the antibody signal’s location. If a distinct imaged pattern does NOT align with...literature and is unrelated to the target protein, we categorize the antibody as having non-specific binding in...

  9. Tips for Technical Support Calls

    Type
    Blog Post
    ... biological reagents have a ton of data on the catalog page. It’s not always super easy to navigate the...originally described in a peer-reviewed scientific publication. We love science, but we don’t have the resources...questions about an Abcam antibody. Have your catalog number ready. I've attempted to answer several ...Addgene support, make sure to mention the plasmid catalog number, or at least the plasmid name in your initial...be fairly fast, getting the full answer to a complicated question may take some time. If the customer ...message Answering customer service questions is a delicate balance of knowledge, patience, and diplomacy,...

  10. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ...chemical replication of RNA in the absence of evolved enzymatic machinery (i.e. RNA replication with no...Guangzhou and the Science City The conference was located in the modern, relatively new “Science City” area...and technology companies and start-ups choose to locate there. I made time to go into the city of Guangzhou...inheritance that might be less error prone than DNA replication. He too mentioned how “surprising” small RNAs...Steitz quipped that RNA in the ribosome is in a complicated conformation “not like DNA which has a boring...

  11. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    ...targeting anywhere in an exon. But things get more complicated if you’d like to use homology directed repair...They created a selection phage (SP) to carry catalytically dead SpCas9 (dCas9) fused to the bacterial polymerase...test xCas9 3.0-3.13, Hu et al. first restored catalytic residues D10 and H840. To ensure that the clones...loci displayed a broad range of PAM sequences, indicating they may be less predictable than SpCas9 off-... that we have only scratched the surface of modifications that can be made to Cas9. It is a pleasant surprise...editing rates. This PAM flexibility should expand applications that require very specific targeting, like homology...

  12. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...so most labs opt to purchase adenoviruses from dedicated adenoviral production labs or companies. Production...something you may have to optimize for your specific application. With mammalian systems, it is standard to do...more cells, while AAV can be titrated following purification using qPCR or digital droplet PCR. Insect cells...to collect your viral supernatant. Baculovirus replicates inside insect cells, and its lifecycle ends in...titer the virus, or go straight into your viral application, like protein production. Titration of baculovirus...Sf9 Time to collection 2–4 days 5–10 days Purification Ultracentrifugation (AAV); centrifugation (lenti...

  13. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    ...antiviral defense). Cascade is brought to the target location using a gRNA and can then recruit Cas3, which ... type I, the most abundant type in the Class 1 category. These type Is can then be further broken into...of DNA much like a fisherman would reel in a big catch. It was originally suspected that Cas3 moved a couple...Mashimo, T. (2022). Genome editing technology and applications with the type I CRISPR system. Gene and Genome...unidirectional genome editing in human cells. Nature Communications, 10(1). https://doi.org/10.1038/s41467-019-...Koonin, E. V. (2015). An updated evolutionary classification of CRISPR–Cas systems. Nature Reviews Microbiology...

  14. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ...RESCUE in therapeutic applications, the authors tested various dRanCas13b truncations that would allow packaging...longer stretches of duplexed RNA. Using an ADAR2DD catalytic mutant, they showed that editing is mediated by...into adeno-associated viral vectors, they tested truncated ADARDDs to see if they could shrink the construct... to deaminate cytidine and used dRanCas13b (catalytically inactive Cas13) to target the cytosine deaminase...construct for viral delivery. They found C-terminal truncations allowed the same or improved editing ability ...

  15. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ...retroviruses ” (Cullen, 1992) because of their more complicated genomes. Here, we will mostly use the gamma-retroviruses...wild-type viral particle — and of the normal, replicative life cycle — can help you understand these seemingly...of a non-living viral particle, and how we as dedicated researchers can exploit it. This blog was written...Hutcheson for her assistance with lentiviral replication.  Resources and References More resources on...101: Types of Viruses Viral Vectors 101: Viral Applications Resources on addgene.org Addgene's Lentiviral...acid of the Bryan strain of Rous sarcoma virus: purification of the virus and isolation of the nucleic acid...

  16. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ... Aptamer biosensors are used in a variety of applications, including detection of environmental pollutants... opens aptamers for use in many entirely new applications including aptamer based fluorescent biosensors... render this approach less sensitive in most applications. Learn About Fluorescent Biosensors Light-up..., MG proved to be too toxic for many in vivo applications, but this work proved the concept that RNA could...and biochips. Ultimately, there are just some applications that lend themselves to aptamer-based instead...Kyung-Mi, et al. “Aptamers and Their Biological Applications.” Sensors (Basel). 2012; 12(1): 612–631. PubMed...

  17. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ...a protein’s stability or post-translational modifications, or study how a target protein interacts with...Protein A and Protein G are not suitable for all applications, such as protein isolation from serum. In this...capture antibody interaction and/or the downstream application. Typically, if the target protein will be detected...and an isotype control antibody. Alternative applications Oftentimes, an IP-appropriate antibody simply...identify DNA binding proteins involved in histone modification. Similarly, RNA IP (RIP) can be used to isolate...resources References:  Björck L, Kronvall G (1984) Purification and some properties of streptococcal protein...

  18. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ... they are based on light which is very easily scattered by tissue. Ultrasound, on the other hand, is a...protein nanostructures called gas vesicles, which scatter sound waves and thereby produce ultrasound contrast...limiting their applicability. They also did not scatter ultrasound nonlinearly, making them difficult to...out to develop next-generation bARGs that could scatter ultrasound nonlinearly and be easily expressed ...promoter pBAD and a low-copy number origin of replication (p15A) worked best under a wide range of inducer...smaller genetic footprint, and nonlinear acoustic scattering. The best candidate for this was the Anabaena...

  19. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    ...Red arrows indicate a conversion directly catalyzed by the base editor. Black arrows indicate natural cellular...researchers can now select the best tool for their application from among many options.  Conclusion With these...engineered human uracil DNA glycosylase. Nature Communications, 15(1), 4897. https://doi.org/10.1038/s41467...via engineered uracil-DNA glycosylase. Nature Communications, 15(1), 6397. https://doi.org/10.1038/s41467...

  20. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...positively charged polycation, polybrene. Interestingly, one study found that alternative polycations such as DEAE-dextran... freshly collected lentivirus for downstream applications. While this ensures that the highest titer virus...lentivirus may want to consider testing a variety of cation and serum sources to ensure the best possible transduction...establishes the best conditions for specific applications, stick with them! Changes in such variables ...efficiency of lentiviral vectors: effect of sera and polycations.” Molecular Biotechnology 53.3 (2013). 308-14...

  21. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    ...EthoVision. Consult the literature to find all the applications of optogenetics relevant to your field. Ultimately...Inhibition of neuronal activation is a little more complicated since basal neuronal activity in your brain region...technology is improving and may one day reach the sophistication of mouse transgenics. Everything that has been... and memory in opioid addiction and science communication/science writing. Derek is also a pianist and...rat lines: tools, techniques, and optogenetic application to dopamine-mediated reinforcement." Neuron 72.5...sensitivity optical neural silencer: development and application to optogenetic control of non-human primate cortex...

  22. 5 Science Rap Videos & Their Creator: Tom McFadden

    Type
    Blog Post
    ...advice on how to dive into new forms of science communication. Click here to listen to the interview with ... from Addgene.      Check out more science education tools like Verse Two and Fill in the Planck at...with us. Tom McFadden is a YouTuber and science educator looking for creative ways to implement the next...next generation science standards and communicate science. Learn more about Tom on his youtube channel ...Community Check out All of Our Podcasts Find Great Educational Content in our Plasmids 101 Series Additional...

  23. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    ...variety of species, cell types, and research applications. Labeling neurons with Brainbow The Lichtman... Brainbow 3.1 and 3.2 (described in the same publication) are unique because by default, they are not ...not fluorescent. Instead, they incorporate a truncated, non-fluorescent form of YFP as the default state...recombination can be detected by immunostaining of the truncated YFP. In Thy1-Brainbow mice, some neurons expressed...level of these XFP is not strong enough for all applications. To resolve this, similar systems have been ...transgene in an adult fly. As a stop cassette is located directly upstream of the fluorescent protein genes...

  24. Replacing Paper: Tips for Choosing an Electronic lab Notebook

    Type
    Blog Post
    ...priorities in most cases include IP protection, publications and funding. If any of these three pillars crashes...budgets, “a large percentage of survey respondents indicated that cost was a significant barrier to ELN adoption...to digital for example): “… 99% of respondents indicated that ease of use would influence their ELN choice...that their research ‘doesn’t fit neatly into one category;”. The study also showed that It is important ...to keep all their data in one place and avoid scattering intellectual property across various software...collaborative functionalities include comments, notifications, sharing files, assigning team members to certain...

  25. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...studying genetic modification for their ease of manipulation and their ability to be replicated. Because E...., 2004). Additionally, breakthroughs in gene modification tools and methods such as CRISPR have vastly...Escherichia coli as a Model Organism and Its Application in Biotechnology. In Escherichia coli - Recent...Physiology, Pathogenesis and Biotechnological Applications. InTech. https://doi.org/10.5772/67306 Koornneef... bacteria with superior performance. Nature Communications, 9(1). https://doi.org/10.1038/s41467-018-04899...

  26. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ...that the oversized AAV genomes are fragmented or truncated when packaged and then following transduction,...and may need to be optimized for each unique application (Trapani et al., 2015).  Expression of smaller...transgene to help prevent the accumulation of such truncated proteins. Examples of use:  Ghosh et al., 2008...design, donor A may be capable of expressing a truncated protein. This can be avoided by excluding a stop...JS (2011) Gene Therapy of mdx Mice With Large Truncated Dystrophins Generated by Recombination Using rAAV6...Improved dual AAV vectors with reduced expression of truncated proteins are safe and effective in the retina ...

  27. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...variety of small RNA types to choose from and can be catered for your specific experiment. For this blog, we...is an umbrella term for a number of different applications that involve different types of small RNA molecules... by a ribonuclease called Dicer. In a great simplification of the process, Dicer chops off the loop of...sequence is highlighted in green, with red boxes indicating mature siRNA. Hairpin loop is in between the ...sequence is highlighted in green, with red boxes indicating mature miRNA. Hairpin loop is in between the ...for your research arsenal, and there is still a dedicated field of study for improving these tools. Next...

  28. Evolution of Lab Techniques

    Type
    Blog Post
    ...comparison to a control sample. Southern blotting applications range from confirming the results of cloning...interest by recording DNA abundance throughout the amplification process, rather than just at the end as in standard...experiments allowed researchers to identify the location of genes. With this knowledge, researchers were...was added and when. Compiling the identity and location of a given nucleotide, a researcher (or a simple...sequence thousands of reads at the same time. Other applications of sequencing, such as confirming the results...replaced these, these techniques still have useful applications and have informed many current techniques. For...

  29. Addgene's Website Update

    Type
    Blog Post
    ...and easily navigate to educational resources, tools, collections in our catalog, and the Addgene Help ...search bar. And if you’re in the mood to browse our catalog, you can use the links under the search box to ...scrolling up and down the page.  Figure 3: The Education & Tools menu Further down What happens if you ... mission, links to our deposit, request, and educational resources page, and previews for our three most...

  30. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ...site, or when using chemogenetic tools, where modification of cellular activity outside of your experiment.... A. (2023). Various AAV Serotypes and Their Applications in Gene Therapy: An Overview. Cells, 12(5), ...comparative analysis of constitutive promoters located in adeno-associated viral vectors. PLoS ONE, 9(...Approaches and Further Perspectives of AAV Tropism Modification. Pharmaceutics, 13(5), 750. https://doi.org/...Retrograde Serotype Viral Vectors 101: Viral Applications...

  31. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ...channelrhodopsin-2 (ChR2), a blue light-activated cation channel, from the archaebacteria Chlamydomonas ...stimulation. GPCR signaling GPCR signaling is a complicated process involving many downstream signaling mechanisms...break your experiments!  First, closely read the publication, including the supplemental data. You’re most... both of which have extensive collections of publications and information on hardware, optogenetics tools...2003). Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proceedings of the ...bidirectional dual-color control of neurons. Nature Communications, 12(1), 1–20. https://doi.org/10.1038/s41467...

  32. Is this the right place for me? 8 tactics for choosing a lab

    Type
    Blog Post
    ...honestly evaluate your own style of work and communication before making this important decision. I’ll ...Prepare by going sleuthing on the lab website, publication records, LinkedIn, ResearchGate or Twitter. Be...the appropriate first authorship position on a publication of her project and made another lab member first...– including failure to acknowledge good work, allocation of meaningless tasks, removal of responsibility...connections such as serving on Boards? Does the publication record show evidence of industry collaboration...skills guide Learn about careers in science communication Read posts on mentoring for scientists Resources...

  33. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...piggyBac™ technology can be used for numerous applications, including transgenesis, gene-trap screens, ...of the piggyBac™  transposase/transposon gene modification system and the scarless, Footprint-Free™ removal...technology that may be used for a variety of research applications, including making transgenic cell lines and ...PMCID: PMC2567143. 6. S. Radecke, F. Radecke, T. Cathomen, and K. Schwarz, “Zinc-finger nuclease-induced...oligodeoxynucleotides: wanted and unwanted target locus modifications.,” Mol. Ther., vol. 18, no. 4, pp. 743–53, Apr...Addgene.org Find Vectors with the piggyBac™ Transposon Catch up on Everything CRISPR with Our CRISPR Guide Deposit...

  34. Developing Transferable Skills During Science Training

    Type
    Blog Post
    ...speak for you (and you for them) Communication Being able to communicate effectively is part of what makes...you can. The colleagues you help will be future advocates and references. Collaborate with another lab,...productive. Take all the opportunities you can to be communicating about science with others. If opportunities ... (be on a speaker committee) Follow industry publications like FierceBiotech to “learn the business” Attend...

  35. Why Do I Need an MTA Anyway?

    Type
    Blog Post
    ...results, modifications, and inventions created by the Recipient Right to file patent applications on modifications... about intellectual property rights, pending publications, and commercial use often come up. An MTA can...trials Acknowledgement in recipient’s future publications relating to the material Limited liability...

  36. Searchable and Sortable gRNAs for Your Next CRISPR Experiment

    Type
    Blog Post
    ...with the gRNA), application, cas9 species, PubMed ID, and depositor. The CRISPR applications we currently...shown to work in CRISPR experiments. Addgene’s dedication to resource sharing has led us to develop a searchable...The efficacy of gRNAs is impacted by the target location, target sequence, and model system. Even if a ...and final experimental outcome in the original publication. As you develop and confirm new gRNAs, please...

  37. Seeing Red: Simple GFP Photoconversion

    Type
    Blog Post
    ...Since the first research applications of GFP were published in the 1990s, biologists have spent a lot...color was observed after excitation at 488 nm, indicating that spontaneous conversion did not occur. Tobacco... Sattarzadeh et al., Sci Rep. 2015 CC BY 4.0 Applications and limitations of GFP photoconversion One concern...2):131-3.  PubMed. Find plasmids from this publication at Addgene. Subach OM, Patterson GH, Ting LM...nmeth.1664. PubMed. Find plasmids from this publication at Addgene.   Addgene’s Fluorescent Protein...

  38. Who Gives a Tweet? 9 Facts About Scientists on Twitter

    Type
    Blog Post
    ...mostly because of this article: “A systematic identification and analysis of scientists on Twitter.” This...from the United States Standard Occupational Classification System  and Wikipedia. 2. Social and computer...: Portion of Tweets with Scientific Domains, Categorized by Discipline. Biologists, neuroscientists, and... Y., & Sugimoto, C. R. (2017). A systematic identification and analysis of scientists on Twitter. Plos...Scientists 5 Reasons to Use Reddit for Science Communication 10 Ways to Share Your Science Why Scientists...

  39. Working 9-5: an Addgene Co-op

    Type
    Blog Post
    ...working in your chosen field. This cooperative education experience (co-op) is what has brought me, a second... QC/QA team as a DNA Sequencing Intern. The application process When I first saw the comprehensive job...for the intended position. After submitting my application, I was contacted to participate in a brief fifteen...the depositing labs intended. With the use of applications like SnapGene and Geneious, I was able to determine...currently working on projects for the Science Communication function at Addgene. Working a 9-5 As it turned...

  40. Celebrating Outstanding CRISPR/Cas9 Achievements at the Dr. Paul Janssen Award Dinner

    Type
    Blog Post
    ...adaptive bacterial immunity and its potential applications in rewriting the genome. Their award acceptance... same Polish sub-dialect making their Skype communications extra fun) who shared first authorship on their...simpler 1 protein - 1 RNA system.  Dr. Charpentier advocated for the crucial role of basic research in the ... area will accelerate the launch of clinical applications. During the panel discussion, Dr. Mello took...for billions of years (…) Bacteria are really sophisticated”.     New to CRISPR/Cas9 Technology? Learn ...
Showing: 481 - 520 of 1023 results