...contributed by our depositors, Addgene strives to make it easier for you to find the plasmids and publications...and the original depositor. Our citations feature makes it so that, when you cite a plasmid from Addgene...who may be working on similar projects and should make it easier for them to find your work later. We also...process. Special collections pages In an effort to make it easier for you to browse through the many plasmids...

...collection, the fluorescent proteins are, which makes mixing and matching a relatively easy proposition...Davidson lab for the last four years. “Researchers can make their own target if it is not already made for them...For more information on choosing a backbone and making fusions, see this blog post from Joachim Goedhart...Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection.  mEmerald plasmids and many...

...mapping These tools are for viewing, editing or making plasmid maps, but can also analyze and annotate...also has a suite of molecular biology tools and can make plasmid maps. Free for academic users. Serial Cloner... blank spaces, from the text of a DNA sequence, making it ready to copy and paste into other applications... Addgene blog: Learn how the CRISPR software matchmaker can help you choose the best CRISPR software ...

...video. Don't worry - no pipettes were harmed in the making of this photo!  Here, I was also given a mixture... understand the importance of using analytics to make decisions on content strategy. I really believe ...different best practices, create an email strategy, and make the occasional TikTok. I was also responsible for... my time here comes to an end, my worries about making connections with coworkers seem, in retrospect,...

...adopting a few meaningful changes in the lab could make a big difference in our global impact as scientists... environmental impact, but we can all do more to make research more environmentally friendly. Adopt one...Universities and companies have the potential to make a huge impact directly on waste, but also on culture...bring ideas to your workplace and you could end up making a huge impact on how waste is handled. A lot of...

...one of the most challenging [parts of my job], so making it a success feels like a real accomplishment.”... to take over hosting them. Since then, webinars make up a larger part of Hans’ workload. “I am currently....” A few other tips: “Learn HTML, write a lot to make sure you don’t hate it, and be flexible. The best...could be featured in future career blog posts. And make sure to subscribe to the Career Channel of the blog...

...medical use. Find More Optogenetic Tools Like pDawn  Making cassava plants resistant to cassava mosaic virus...this drought tolerant plant, Dr. Robertson aims to make use of the CRISPR/Cas9 technology to engineer plants...for example this yeast cloning kit) she intends to make yeast capable of desaturating fatty acids. Identification...Associate Professor (above, far right), thrives to make affordable biosensors that detect herbicides, such...

... in person to figure out ways we can make Addgene better and make scientists’ lives easier. Why transition... moved into this position because I want to help make the research enterprise work better and because ...learn ways Addgene can improve in its mission to make research easier. I’m hoping my story demonstrates...

... updated Sep 10, 2020 by Jennifer Tsang. CRISPR makes it easy to target multiple loci - a concept called...expressing multiple gRNAs on the same plasmid, you’ll make sure that each cell that gets the plasmid contains...increases the chance that all the edits you want to make will happen.  There are many reasons to multiplex...constructs with multiple promoter-gRNA cassettes, making them advantageous for small capacity vectors like... mammalian option described below, plasmids for making polycistronic gRNAs are also available from the...

... the absence of guide RNA, preventing Cas9 from making off-target cuts.   Pro tip! X-ray crystallography...base. In Figure 5, we can see why: two arginines make base-specific hydrogen bonds to the guanines, but.... Residues R1333 and R1335 from the CTD (yellow) make base-specific hydrogen bonds to guanines of the ...The HNH domain, which cuts the target DNA strand, makes few contacts with the rest of Cas9 and is highly...BioRender.com.   The exact conformational changes that make Cas9 such a valuable laboratory tool are still an...

...for imaging a protein’s entire life-cycle. This makes it nearly impossible for scientists to use GFP to...large, dim, and permanently attached to a protein, making them hard to amplify and image for long periods...in vivo. These probes however can be expensive to make, hard to deliver into living systems, and often ... proven to fold and function in living cells. To make these scaffolds specific to different targets, the...

...problems through good software design. But what makes software good? Fortunately, that question has been... task, we envision what the user wants to do and make the path to accomplishing their goals as intuitive...the most important controls front and center, and make specialized controls available when needed. SnapGene...For a coding sequence feature that may be used to make fusion genes, detection needs to occur even if one...

...you believe your idea has potential to make a difference, then make sure you’re able to answer every possible... Start with your network You've spent your life making connections and finding mentors -- well, it's time...discuss each project and business decision before making it, to have an internal review system.  On top ...

...method limits screening to a few hundred worms, making the detection of rare events difficult. Dickinson...resistance and the Roller movement phenotype. To make these markers easy to remove after injection, they... to generate a self-excising cassette (SEC). To make it possible fluorescently label proteins in C. elegans...fluorescent protein fusions, this system could be used to make many other genome modifications, including targeted...

..., as it can edit near to or far from PAM sites, making it less constrained by PAM availability than other...within the base editing window, while prime editors make a specific edit defined by the pegRNA. In cases ...the non-edited strand, they can cause the cell to remake that strand using the edited strand as the template... These variants are worth testing if you want to make your prime editing as efficient as possible. ...Some examples include: TwinPE: two prime editors make complementary edits to opposite strands of the DNA...

...pombe (fission) and S. cerevisiae (budding). Yeast make for an excellent model organism for a few reasons...phaffii is highly efficient at protein secretion, making harvesting recombinant proteins a cinch. This is...it doesn’t secrete its own endogenous proteins, making the recombinant protein its major secretion. Beyond... removing methanol from the media. This feature makes the strain tractable for peroxisomal studies on ...

...are able to make when they have access to shared materials, and then pay it forward by making the new materials...has been able to rapidly propagate this strain, making it available to scientists around the world. This...research by removing logistic hurdles to continue making materials available rapidly to scientists (Nature...

...generating/replenishing kits, is to find new ways to make lab processes more efficient. Being in the Boston...there because both LabCentral and Addgene aim to make scientific advancement easier. NEBNext Ultra FS ...we did at the workshop (the kit is mostly used to make new libraries as opposed to replicating them like...new without spending a lot of time developing and making space for the techniques ourselves. As you probably...

... strategies can make socially distanced work more creative. An example could be making lab meetings more...without leaving Zoom. The annotate function can also make the screen-sharing experience more interactive. ...: “Science is increasingly done in large teams, making it more likely that papers will be written by several...

...Christopher Voigt and Jeffrey Tabor have been working on making E. coli light responsive since 2005, when Tabor...systems continue to be optimized by Tabor’s lab, making light-controlled gene expression in E. coli easier...the response regulator encoded in the genome. To make the system less bulky and easier to use with other...Schmidl et al. chose to optimize these systems to make them more tunable and user-friendly. They experimented...

...the earliest) or a functional one. Pick one that makes sense for your skills, and the job you are applying...seconds on a resume when they first look at it. Don’t make them have to guess what you are capable of. You ...might know a thing or two about computer modeling. Make sure to focus on these skills. The only exception... yet for that dream job. Also, many universities make their job postings and salary expectations public...

... digest tips and tricks: The following tips will make it easier for you to obtain a useful and informative... recognition sequence. Most enzyme manufacturers make High Fidelity versions of the endonucleases and/...before loading them. The glycerol in the buffer will make sure your sample settles in the gel well and the...wrong! Quantify your DNA. Loading too much DNA will make it difficult to obtain crisp bands and analyze the...

...think. So, if you’ve ever wondered what goes into ‘making virus’ or what gag and pol stand for, we demystify...recombinant adenovirus is missing the endogenous E1 gene, making it replication deficient. This is typically supplied... some of the plasmids in our collection. Want to make a vector construct on your own? No problem, we are...discussed are compiled for reference below. Happy virus making!  References and Resources Kulanayake, S., Tikoo...

...value. Types of biosensors So now that we know what makes up a biosensor, we can discuss what flavors of sensors...difficult to achieve in whole organisms, but AAVs make it possible. In AAV systems, different serotypes...expression and downstream monitoring. Nervous about making a virus for your biosensor? Don’t be! We have you...viral resources and safety tips. Still worried? We make virus with some of our most popular biosensors already...

...variety of educational resources in an effort to make scientific knowledge broadly available.  The ability... return the matching resources in an order that makes sense to the user. The devil is in the details of... of course: what does “best” mean? and what “makes sense” to a user? Moreover, there are many ways a user...institutions. There are many more technical details that make our new search engine unique. Most importantly, ...

...efficiency. Check out Addgene’s blog to learn about making your own competent cells and our protocols page... encodes a green fluorescent protein gene which makes the bacterium fluoresce green. Comic by Maya Kostman...Lytic phages hijack the bacterial hosts machinery to make more viral particles. Eventually the cell lyses ...bacterial chromosome and allows the donor bacterium to make a small “bridge” or sex pilus that attaches to the...

... explore small-scale opportunities where I could make mistakes and learn from them. I started at my previous...Beyond Academia: Stories and Strategies for PhDs Making the Leap to Industry." He successfully transitioned...Resources  Resources for aspiring managers Books: The Making of a Manager by Julie Zhuo Radical Candor by Kim...Managing vs. Leading Management for Scientists: What Makes a Good Leader Anyways?  Transferable Skills Guide...

... task will keep you from repeating your work or making similar mistakes in the future. Stay organized...camelCaseFormat since spaces between each word can make it difficult to accurately call a file. Version...your exact changes. Alternatively, you can simply make a note in your methods section about the edit that... Your Experiments Try out the CRISPR Software Matchmaker Additional Resources on Addgene.org Find Protocols...

...will be a positive asset throughout your career? 1. Make scicomm practice a priority The good news for young...of the work you presented may also be unclear. 2. Make the audience your primary focus When writing a paper...address that 'why' question effectively, you need to make your audience the focus of every sentence you write...know this? How will they react to it - what will it make them think? Does this lead to the conclusions I ...

...sometimes called “VUS”) function is unknown. How can we make a decision for each patient if the majority of information...high-throughput manner and therefore push us closer to making cancer precision medicine a reality. Challenges...have worked closely with our Addgene colleagues to make these available to the scientific community. Functional... Institute of Harvard and MIT and is focused on making precision functional genomics a reality for future...

...Zetsche, B., Gootenberg, J., Abudayyeh, O., Slaymaker, I., Makarova, K., Essletzbichler, P.,  Zhang, F. (2015... diagram below for an example of how this works. Make sure to order 5’ phosphorylated oligos or treat ...single plasmids are generally more efficient. Pro-Tip Make sure to reverse the orientation of the crRNA’s direct...

...feature, you can compare different Cas enzymes to make sure you are getting the desired features and cutting...and discover orthologs with similar properties (Makarova et al). CasPEDIA is an evolving database, and... to it! The CasPEDIA Consortium will, of course, make efforts to keep it updated with new entries and ...Structural Biology. https://doi.org/10.32942/X2C31F     Makarova, K. S., Wolf, Y. I., Iranzo, J., et al. (2019)...

...summaries of recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you...Parton Lab recently created a flexible system that makes use of pre-existing transgenic lines, termed BLITZ...system provides researchers better flexibility and makes the approach more accessible, as it reduces the ... be packaged into an AAV system. dCas9-SSAP will make the editing of kilobase-scaled sequences more efficient...

...character you painted in your personal statement. Make certain to choose your recommenders based on your...individuals are inundated with requests. The easier you make it for them, the likelier you will end up with a...lighting for a smooth performance. Most importantly, make certain that everyone you live with is aware of ...consistent with your personal statement, while you make the ultimate case: why you? 7. Strength in numbers...

...behind. While commercial kits are available for making small point mutations, large deletions/insertions...Current SDM methods rely on in vitro assembly. To make a point mutation, you design two oligonucleotide...instead of relying on in vitro assembly, they could make the E. coli do the work by using the principles ...-12 kb range. REPLACR-mutagenesis also requires making your own electrocompetent cells in which the phage...

...between blue and white colonies. Take care in making your plates: X-gal is light and temperature sensitive...autoclaving. If spread on top of pre-made plates, make sure it is evenly distributed and allow sufficient...specific analysis, like PCR or restriction digest. Make sure you use a proper E. coli strain (i.e. contains...JM109, STBL4, JM110, and Top10 are a few examples. Make sure you use a proper plasmid (i.e. contains the...

...how the new kit works, the research the kit now makes possible, and how it compares to his CRISPR/Cas9...step. The kit also has other unique aspects that make it especially well-suited for use in stem cells....hands of pretty much all investigators. Our kit makes it extremely simple to design TALENs. It only takes...apples-to-apples comparison of all those mutations. If you make induced pluripotent stem (iPS) cells from patients...

...scientists commonly rely on RNA isolation kits to make their life easier. Recently, we published a blog...cold and work quickly to mitigate RNA degradation. Make sure to use DEPC-treated or RNAse free water. If..., 2006).  View our protocol for RNA extraction! Making the simple protocol even more foolproof (still ... if you need really clean RNA, it’s important to make sure that when extracting, to only take the aqueous...

...experiments, and here we will focus on a unique gene that makes the popular GatewayTM method possible: ccdB. But... more about it? Read on to find out how ccdB can make your cloning experiments a little easier. One of...contain your insert of interest. Simply stated, ccdB makes cloning easier by selecting against vectors that...not contain the recombinant plasmid and therefore makes the cloning process much more efficient, as one ...

...properties of the fluorophores you are considering to make sure that they’ll be compatible with your instruments...properties allow for efficient signal amplification, making biotin conjugated antibodies good options for detecting...antibody. 2) Endogenous cysteines, such as those that make up the disulfide bonds between antibody chains. ...antibodies yourself, there are kits available to make the process quick and easy. Before getting started...

... see who you are when you look at their profile. Make your professional contact information available.... be important for a good friend, but if you are making a professional inquiry, you always want to read...type of connection can help you gather information, make introductions, and give advice for your interviews...should be as good or better than your resume Don’t make the mistake of having a shoddy profile and not updating...

...address the research challenge. Now is the time to make bold statements. While some of the statements may...6-12 months to give feedback on the application. Make sure to invest time and consider the reviewer's ...the next application. Although some comments may make it clear that the reviewer misunderstood the point...that was being made, take it as an opportunity to make clearer statements on the next round of applications...

...or a G to A change on the opposite strand). ABEs make an A to G change (or a T to C change on the opposite...catalytically dead or nickase) fused to a deaminase that makes the edit A gRNA targeting Cas to a specific locus...efficiency for mammalian edits is to ensure the editors make it into the nucleus and that they are expressed ... 98-99% target modification in primary T cells, making them a promising tool for cell therapy applications.... J., Murray, R., Packer, M. S., Rinaldi, C., Slaymaker, I. M., Yen, J., Young, L. E., & Ciaramella, G...

...changes (Figure 1), but it can be challenging to make these base changes, especially in primary human ....20-m to edit primary human T cells efficiently makes it a promising tool for T cell therapy development...robust editing in human cell lines and primary cells makes them a great addition to the base editing toolbox..., Liquori AJ, Murray R, Packer MS, Rinaldi C, Slaymaker IM, Yen J, Young LE, Ciaramella G (2020) Directed...

...Tweeting and texting, actually writing an e-mail or making a phone call is a big commitment when you’re busy... to consider before you commit: Double check to make sure the answer to your question didn’t come with...expedite and improve your customer service experience: Make sure you’re contacting the correct company. This...as possible about the issue. For Addgene support, make sure to mention the plasmid catalog number, or at...

...for antibodies. Recombinant antibody technology makes it easy for scientists to share, remix, store, and...     How are Recombinant Antibodies Made? To make recombinant antibodies, you first need to know the...  Second, plasmids are easy to store and share, making recombinant antibodies the most practical choice... encoding antibodies, scientists will be able to make higher affinity antibodies, modify antibody function...

...limiting reagent for my experiments, so I always make more AAV than I think I need. Unlike lentivirus,.... If you’re new to creating gradients, practice making the iodixanol layers before doing your first purification...purified and buffer exchanged virus, remember to make one or two small aliquots to use for qPCR titering...Additional resources on the Addgene blog Find out what makes AAV a versatile tool for gene expression in mammals...

...Jacob Corn contacted us in 2014 to ask if we could make a model of Cas9 based on the structure published...Jennifer Doudna’s new book A Crack in Creation. Making models more useful through flexibility It is common...the several Lys/Ala mutations reported by Ian Slaymaker and colleagues to reduce off-target hits in a ...our hands-on teaching tools that are designed to make the molecular world real for students. Many thanks...

... jobs needed to be done (Who is responsible for making the running buffer this week?). Scientists negotiate...were hoping that your lab mate would have time to make comments on your manuscript. If you give her this...practice and hone your skills. This practice will make negotiating feel more natural. Creative negotiation...the person/team for help? What can I offer them to make it a win-win situation instead of just extra work...

...to help make the most of these pages, we've written this post to guide you through them and make the best...can be found in this location as well. These maps make it easy for scientists to see if the plasmid would...their plasmids are used in a publication. Addgene makes this easy by providing a template citation for you...