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Showing: 521 - 540 of 1088 results
  1. Summer SciComm: Preprints

    Type
    Blog Post
    ...As a scientist, perhaps the most important part of your research is communicating the results of the ...the work. Yes, those control experiments are important — but without letting other scientists know the results...conference poster or talk, or submitting patent or grant applications; to informal settings like lab meetings...amounts of access or feedback. One of the most significant shifts in scientific communication is the rising... of listing “Submitted” articles in their CV or grant applications. A number of organizations (such as... or invite attendees to read and comment online! Want the general public (say, policymakers or community...research materials available to the community significantly sooner. Plus, authors can include the Addgene...
  2. Viral Vectors 101: An Introduction to AAV

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    Blog Post
    ...optogenetics, and chemogenetics experiments. Recombinant AAVs (rAAVs) The primary application of AAV is...the deliverable DNA) are removed, resulting in recombinant AAVs (rAAVs) with maximum “trunk” space for genetic...specific cells and/or tissues is one of its biggest advantages. Specificity can be achieved using specialized...specificity; this is particularly helpful when wanting to target specific neuron subtypes, since AAVs ...and our AAV data hub to see if others have used relevant viruses for your system. This post was contributed...and Richard J. Samulski. "Self-complementary recombinant adeno-associated virus (scAAV) vectors promote..."Development and optimization of a real-time quantitative PCR-based method for the titration of AAV-2 ...
  3. Career Coaching for Scientists: Why and Where Do I Find One?

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    Blog Post
    ...White and Anthony Collmann of White Consulting frequently do sessions by Skype or phone. Samantha Sutton ... goals are for the coaching relationship. Do you want to change jobs? Grow your professional presence?... consultation meeting or call for free.  Take advantage of this to get the information you need to choose...a scientist from academia to industry, you might want to consult the Cheeky Scientist. Their motto is ...coaches I have mentioned. I have seen a lot of fantastic online content for PhDs and postdocs posted by...bottom line – there are lots of people out there who want to help. Don’t be afraid to ask and to make an investment...
  4. Lab Automation at Addgene

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    Blog Post
    ...machine or robot out there that does exactly what you want plus way more than you thought it could ever do!... taking a lot of time. Enter the Biomek FXP! We wanted a machine that would improve our transformation...purification We're developing ways to prep large quantities of DNA because some countries just can’t receive...fills all those vials? Well, it used to be a lab assistant, but with increased plasmid demand it was too ...fill 400 vials in 35 minutes compared to a lab assisstant filling about 200 vials in 45 minutes by hand...have used in the lab. It’s especially useful if you want to avoid sore fingers. There are other process ...about more lab updates as we develop them. In the meantime, if you have any great lab automation tips and...
  5. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

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    Blog Post
    ...process involves a significant amount of QC. First, we perform Picogreen quantification on each and every...high-quality isolated DNA samples of sufficient quantity for sequencing. This process is completed in a... preparation process and includes Picogreen quantification of various intermediate steps, a sizing electrophoretic...electrophoretic gel, and qPCR quantification of the final libraries. A broader view of the whole plasmid... Analysis Platform to Assess the Identity of Recombinant Adeno-Associated Viral Preparations from Viral...
  6. Plasmids 101: Modular Cloning

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    Blog Post
    ...modular cloning toolkit for genome editing in plants. BMC Plant Biol 20, 179 (2020). https://doi.org/10.1186...libraries depend on many gRNAs delivered with a relevant Cas nuclease. MoClo can be used to assemble multiplexed...are not available to those working with yeast or plants. Using MoClo, or better yet, a pre-made MoClo kit...achieve a desired effect. This is commonly used in plant biology to achieve traits such as disease resistance...blog on choosing a MoClo strategy and kit if you want to learn more about this cloning technique! Many...Cloning Guide EcoFlex MoClo Kit CIDAR MoClo Kit MoClo Plant Parts Kit Resources on the Addgene blog High Complexity...
  7. Viral Vectors 101: Chemogenetics

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    Blog Post
    ...equipped using fast-acting optogenetic tools. The advantages provided by chemogenetics are instead that it...Targeted and specific delivery becomes especially important for studies in neuroscience where specific tissues...neuron need to be targeted. Targeting is also important in the development of therapies when expression...Picking Ligands Picking the right ligand is also important for building a chemogenetic system. Your experiment...cellular response you aim to manipulate. It is important to consider that the same receptor can elicit ...drug that only binds to one receptor, so it is important to fully characterize the compound, or compounds...will promote hyperpolarization. PSAM ligands The advantage of PSAMs lies in their ability to be specifically...
  8. 5 Tips for Troubleshooting Viral Transductions

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    Blog Post
    ...selectable with the antibiotic whose resistance gene is encoded in the viral vector. This antibiotic selection ... 320,000 viruses can infect mammals. Even more abundant are the Earth’s estimated 1031 bacteriophages ...that infect bacteria), many of which are doing important work in our microbiomes. Given that viruses are...tips for your virus experiments. References 1. Anthony, Simon J., et al. "A strategy to estimate unknown...hepatitis B virus DNA and hepatitis C virus RNA quantification as measured with branched-DNA technology." ...PMC84922. 5. Ugai, Hideyo, et al. "Stability of a recombinant adenoviral vector: optimization of conditions...
  9. Plasmids 101: FLEx Vectors

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    Blog Post
    ...fragment. The availability of both wild-type and mutant loxP sites has allowed scientists to leverage this...site-specific recombination. The FLEx switch takes advantage of the orientation specificity of the site-specific...needs. How does the FLEx switch work? Let’s say you want to design a genetic FLEx switch that turns BFP expression...followed by the mCherry coding sequence in the antisense orientation (Figure 1, top). The entire DNA cassette...the gene you would like to rescue with in the antisense orientation (let’s call the gene “GeneR”). The...that your gene of interest starts in the inverse/antisense "off" position and is flipped to the sense (on...Saunders A, Johnson CA, Sabatini BL. 2012. Novel recombinant adeno-associated viruses for Cre activated and...
  10. How-to: 5 Steps to a Great Panel Discussion

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    Blog Post
    ...smallest program. I don't mean have fancy food or a fantastic view (these are nice if you can afford them). ...performance with room for fortuitous improvisation. You want to make it look effortless to the audience, but ...so her ability to control the event is what is important. Choose 4 panelists (maybe 5) and no more. Too...on the topic at hand. You can use this to the advantage of the program. Start the program by polling the...Timing is everything Many of my suggestions are meant to keep the event on time. Nobody likes a program...postures shift, they are checking out. If you can, plant 1-2 good questions with trusted audience members...section. And what about those audience members that want to take over? It is the moderator's job to politely...
  11. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...or the recombinant antibody can be purified from the supernatant (see our Recombinant Antibody Purification...for Recombinant Antibodies You may also like... Antibody Plasmid Collection Antibodies Blog Posts Antibody...inhibitor cocktail to the supernatant. The supernatant containing the recombinant antibody can be used as-is ...cells to produce recombinant antibody. Protocols... Protocols Transfection for Recombinant Antibodies Transfection for...Antibody Guide General Transfection Protocol Recombinant Antibody Purification Protocol Introduction Transfections...to transfect suspension HEK293 cells with recombinant antibody plasmids using Polyethylenimine Max as a...
  12. Degrading DNA with Cascade-Cas3

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    Blog Post
    ...called Cascade (CRISPR-associated complex for antiviral defense). Cascade is brought to the target location..., with Cas3 systems making up type I, the most abundant type in the Class 1 category. These type Is can....com.   The Cascade complex Cascade is an important component of the CRISPR Cas3 system. Most Cascade...is pulled through the Cas3. This mechanism is important for Cas3’s biggest trick. Cas3 likes to bluff,... pairs, stopping either at boundaries defined by anti-CRISPR proteins or running into other proteins naturally...to its full potential. Hopefully more people will want to utilize Cas3’s degradative power in the future...CRISPR 101: Cas9 vs. The Other Cas(s) CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems INTEGRATE...
  13. Tips to Make the Most of a Scientific Conference

    Type
    Blog Post
    ...first things you might want to know is what their project is. Or maybe you want to introduce yourself ...poster: those that want you to walk them through the entire poster, or those that want to look through themselves...to introduce yourself and what you do. You might want to include your university and your advisor’s name...like “Let me know if you have questions or if you want me to walk you through the poster.” Have a notebook...there. Check out the conference website for any relevant hashtags and handles to help you find other scientists...
  14. Academic vs. Industry Postdocs

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    Blog Post
    ...patients. And, to be honest, I’ve never enjoyed grant-writing. The more I talked to people with industry...interested in the drug discovery process, or you want to end up in an industry job, being immersed in ...on which company you choose. If publishing is important to you, make sure that your project will not involve...pages. In academia, however, writing papers and grants is an inescapable and essential part of being able...’re often required to at least attempt to write grants. If you don’t enjoy or excel at writing, an industry...postdoc will free you of the never-ending cycle of grant-writing that is academic life. Of course, because...contrast industry postdocs at those companies. Do you want a postdoc that’s shorter? Longer? Would you like...
  15. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...PEG precipitation of the HEK293 cell culture supernatant because it’s time consuming to prepare the 40%...virus from the cell pellet alone. If yield is important, or if the viral particles of the AAV serotype... concentrations of iodixanol to separate out contaminants from an impure AAV preparation. 15%, 25%, 40%...titration by qPCR Overview: AAV titering by qPCR quantifies the number of genome-containing viral particles... primer/probe set can be used and samples are quantified by comparing to an AAV standard curve, which ...Universal real-time PCR for the detection and quantification of adeno-associated virus serotype 2-derived...R.A., & Samulski, R.J. (2007). Production of recombinant adeno-associated viral vectors for in vitro and...
  16. Plasmids 101: 5 factors to help you choose the right cloning method

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    Blog Post
    ...would at least be guaranteed a successful result? While there are never any guarantees, some cloning techniques... purposes. We’re here to help with that! Four important factors to consider for any experiment are: ...Efficiency For cloning in particular, another important consideration is: The number of inserts ......Resources” section at the end of the post! Speed: Your grant deadline is approaching or your advisor is demanding...looking for a more streamlined approach, you might want to give Gateway a try. This method, developed by... to the use of toxic ccdB to prevent any non-recombinant vectors from propagating. The ligation-independent...
  17. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    ... is our own immune system, which can recognize antigens on cancerous cells and eliminate them from the...ensues, traditional chemotherapies have taken advantage of the universally-dividing behavior of cancer...approaching the Ras problem by exploiting the different mutant versions of Ras that are most prevalent in human...fluorescent protein fusion studies and the generation of mutant cell lines. Further, for each plasmid in the kit...making the results of any such study as clinically relevant as possible. Connecting the research community...community, will soon expand to include several other important pathways. We invite feedback and contributions...of signaling pathways with potential to confer anticancer drug resistance." Science signaling 7.357 (2014...
  18. Plasmids 101: Multicistronic Vectors

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    Blog Post
    ...of detection; they are useful almost anytime you want to express multiple genes in the same cell. Although...expressed from the same cassette is sometimes advantageous, particularly when only a portion of the plasmid...expression levels between two or more genes is important. How do multicistronic vectors work? Scientists... mRNA. However, some bicistronic vectors take advantage of an element called an Internal Ribosome Entry...bicistronic vectors; however, they do have some disadvantages. These elements are quite large (500-600 bp)...vector.3  2A Peptides To overcome some of the disadvantages of the IRES element, scientists have adapted...PubMed. 5. Scalable signaling mediated by T cell antigen receptor-CD3 ITAMs ensures effective negative selection...
  19. Which Fluorescent Protein Should I Use?

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    Blog Post
    ...derived. For a breakdown of GFP, it's variants, and their relevant mutations, check out Marcy's post from...FPs just keeps getting larger with hundreds of variants available. Read on to familiarize yourself with...emission wavelenght range).  By mutating GFP, the variants blue FP (BFP), cyan FP (CFP), and yellow FP (YFP...calculated as the product of extinction coefficient and quantum yield of the protein, divided by 1000. In many ...temperature). pH Stability: This parameter is important if you are planning to express the FP in acidic...with very comprehensive data sheets on many FP variants Plasmids 101: Green Fluorescent Protein (GFP) ...
  20. A Needle in a Base-Stack: Cas9 Structural Biology

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    Blog Post
    ...it is limited to only 20 bases, or why it’s so important to choose a target sequence with a nearby protospacer-adjacent...unpaired strand in the center of the protein. The repeat:anti-repeat duplex pokes out from the REC lobe and...blue.   The REC lobe binds tightly to the repeat:anti-repeat duplex of the guide RNA, confirming its..., but they improve cutting efficiency and are important for robust activity in cells (Jiang & Doudna, ... loop to produce SuperFi-Cas9, a high-fidelity variant that still retains high cleavage efficiency (Bravo...correct positioning of the non-target DNA strand is important for stabilizing the active conformation of the... and different techniques all bring their own advantages—and blind spots—to these questions. Each new ...
Showing: 521 - 540 of 1088 results