...philosophy Customer service can be a hassle. You finally make it through a 20-step phone tree, only to be left... customer support with self service We strive to make our support team available whenever you need them...not have the chance to stop what you’re doing and make a phone call or write out an email. You need help...resources, we will continually update the Help Center to make sure that, whatever your support needs are, we’ve...ll navigate to articles answering your questions making it easier than ever to get the support you need... regular site, www.addgene.org. We’re excited to make these new features available to the Addgene community...valuable feedback. This is Addgene after all, and making your research easier always comes first. Resources...

...produced in abundance, making them useful for in vivo studies. These attributes make frog eggs some of the...physiological, or developmental characteristics that make them ideal for studying in a laboratory environment...Additionally, its status as a Biosafety Level 1 microbe makes it a perfect candidate for introductory teaching...Iannaccone et al., 2009). Rats are larger than mice making them more useful for certain studies that involve...axolotl has several attractive characteristics that make them an ideal model organism; they are relatively...genetics and developmental patterns are conserved, making them great models for human disease research. One... alike.  Arabidopsis has many of the traits that make an ideal model organism: quick development, a large...

...’s critical to make these decisions in the following order of importance: 1. First make decisions based... Then make decisions based on financial values. All too often, we see early investigators making unnecessary...step by step instructions on creating a budget, making decisions based on that budget, and negotiating...science will define your success. Therefore, it only makes sense to reverse engineer your budgetary needs from...you can eliminate from your budget. Every cut you make signifies that you've made a conscious effort to...dependent on a pathology microtome/cryostat, then making a careful decision on the condition and availability...

... studies, making a meta analysis available to the community on ReFigure. ReFigure also makes it easier...during this period occurs infrequently and rarely makes it into the small selection of data found in full-length...early career researchers, we wondered how we could make the insights gained during this time more visible... on science. ReFigure saves time, knowledge and makes your insights discoverable. Watch this quick video...cell lines/model systems ReFigure allows you to make meaningful collections from a large section of open...paper they’re reading has been linked in a ReFigure making it easy for others to discover your insights on...

...culture towards a more generous and rewarding way of making science.   How we started Crowdfight-COVID19...Confirm the expert’s suitability and availability; 3) Make the contact to start a direct collaboration between...realization that there is a great untapped potential to make science more efficient, open, and collaborative....collaborative. We now want to use this potential to make a difference not just against COVID-19, but also in other...These are transient partnerships where a researcher makes a small but crucial contribution to someone else...We are a community of researchers who believe in making science less of an individual effort and more of...bono. If this sounds appealing, join us and help make science a better place. You will receive opportunities...

...summaries of recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you...with the cells’ endogenous ubiquitin ligase, the Kanemaki lab was able to translate AID to yeast, as well...Nishimura et al., 2009, Natsume et al., 2016).  The Kanemaki lab previously developed a CRISPR/Cas9 based method...essential proteins (Natsume et al., 2016). Now, the Kanemaki lab has created a new toolkit of plasmids for ...Yesbolatova et al., 2019) using CRISPR. This toolkit makes it easy to tag your protein of interest to create...create conditional human cell lines. Lastly, the Kanemaki lab demonstrated that auxinole, a TIR1 inhibitor...such as a distal promoter and an enhancer. What makes LADL unique from previous iterations is the addition...

...rushing, you’re much more likely to make a mistake. When you make a mistake, you’re more likely to be ...and the same goes for your workflow. Next time you make a mistake, instead of silently vowing never to do... physical copy of the protocol when you work and make a checkmark next to each step as you complete it...experimental steps are worth doing, and doing well, so make sure you’re taking time to do them carefully and...reach for spreadsheets, printouts, and even label makers to keep things legible. It is faster to spend more...however, many ways for you to reduce the chances of making an error. Try a variety of things, including (and...

...Alzheimer's - a connection I wouldn’t have thought to make (even if science writer Ed Yong’s book “I Contain... skills you’ve developed in your current work to make new insights in neuroscience. Even if you're currently...expertise and can be difficult for individual labs to make efficiently. For select viral vectors in the repository...repository, you can instead have Addgene make the virus for you. Neuroscientists are particularly interested...experiments. This helped reinforce our goals of making more AAVs and educational content to help researchers...for future experiments and to find tools that will make your transition into the field much easier! If there... suggest a plasmid to us, request that we start making the AAV, or help us produce new educational content...

...as a great first-time mentoring experience. They make mentoring less mysterious and train the participants...Provides a good mechanism for accountability that makes mentoring effective Goal setting is easier with ...example, a group of people who are all interested in making a transition. Schedule your first meeting and get...regularly no matter what, even if one person can’t make it. One of the biggest group mentoring predictors...breakfast once a month. All participants must commit to making the meetings a scheduling priority (and last minute... Set concrete goals with input from the group to make the plan better. Track your goal progress as a group...the series to get more advice on mentoring: What Makes a Good Mentor? Choosing a Good Mentor for Scientists...

...during your current job and note what makes your heart sing, what makes you pull your hair out, your successes...opportunities to pivot and grow along the way. That’s what makes it so wonderful. The most challenging part of jumping... in a government setting: drafting legislation, making rules, and implementing the policy within the jurisdiction...career or a part-time passion, there are many ways to make it work for your time and interests.  Action Items... healthcare systems so I could understand how to make new treatments and technologies accessible to patients...healthcare system? Defining your why is what will make you a competitive candidate for any science policy...your interest to future positions, but will also make your transition from academia to the policy world...

...of the Cas9 nuclease (PDB ID: 4OO8 and 4UN3), Slaymaker et al. hypothesized that Cas9 cutting efficiency...enough to keep off-target sites from being cut. Slaymaker et al reasoned that, if they decreased the positive...the electropositivity of the HNH/RuvC groove, Slaymaker et al. made a variety of alanine substitutions...off-target effects genome-wide. Further work by Slaymaker et al. showed that these mutants are not toxic...SpCas9-HF1, and HypaCas9 should enable researchers to make precise edits in mammalian cells and may decrease...look forward to seeing how the research community makes use of these tools and are excited to see continued...improvements in genome engineering! References 1. Slaymaker, Ian M., et al. "Rationally engineered Cas9 nucleases...

...technology and synthetic biology, the tools he’s making available through Addgene, and where all of it ...CRISPR is so orthogonal and programmable that you can make a very large number of regulators. That basically...have the solution. Our paper shows we can easily make orthogonal variation. The first one worked in each...off-target interactions. That’s relevant if you want to make sure you don’t target anywhere else, but for synthetic... been putting together very large toolboxes and making them available. We’ll also be submitting parts ...you want. The last few years, we’ve really been making tons of parts and characterizing the software to...sophisticated as those that living cells and animals make. That will be an outcome of genetic engineering....

...few things you can do to make the experience less stressful. First, you can make arrangements with those...company’s area of expertise. This has potential to make your audience really sit up and pay attention. Your...demonstrate technical and transferable skills that make you qualified for the position. Highlight the outcomes...improving a process or procedure, solving a problem, or making a key decision. Even deciding to stop pursuing ...4.5 hours, which was a lot of extraversion for me. Make sure to get enough sleep the night before. Have ... look professional  First impressions matter, so make sure yours counts. Dress professionally but still...

..., need to stay active and involved if we wish to make positive change. Below I present perspectives on...science happens when scientists have 1) the courage to make mistakes and 2) the integrity to recognize these...scientific evidence. We need representatives and policymakers who not only believe in the separation of church...-two-new-sciences). This is the approach policy makers take in abusing climate science skepticism to promote...averting a potential climate disaster. However, policy makers use these two fingers of skepticism to defend an...world. Justin Sherman is a Scientific Instrument Maker at Intellectual Ventures Laboratory. Vancouver, ...ongoing and difficult job. I urge other marchers to make this their first step, and continue this work by...

...tips on how to make a successful event at your company. The point of this post isn’t to make you all jealous...are busy with many other work responsibilities, we make team building activities a priority and have organized...though we are hiring!) but to encourage everyone to make an effort to build relationships with their colleagues... it simple This may be an obvious one, but don’t make things too complicated for yourself (especially ...for help My grandma used the old adage “many hands make light work” all the time. She was a wise lady for...will save you time and sanity!    10. Communicate Make sure people know that there is something fun happening...

...Gootenberg JS, Konermann S, Joung J, Slaymaker IM, Cox DBT, Shmakov S, Makarova KS, Semenova E, Minakhin L, Severinov...://doi.org/10.1261/rna.054098.115 Shmakov S, Abudayyeh OO, Makarova KS, Wolf YI, Gootenberg JS, Semenova...Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, Essletzbichler P, Volz SE, Joung J,...

...between different projects, you might not be able to make any reasonable progress on any one of them. That...another. Monitoring task progress. For this, I always make sure I have explicit goals in mind for the endpoints...everything in a Word or google doc that I can comment and make notes on as I progress. I’ve also recently started...world, procrastination can easily prevent you from making progress on a task. For instance, even if I had...over the activation energy barrier that comes with making ANY progress, the task will likely be much easier...easier. Having a formal plan for your day will make this easier - if you know you have dinner plans at 6...pm. Hopefully you’ll find this post useful for making your days, experiments, and projects more manageable...

.... Scientists can make a new line of flies in only 6 weeks compared to months to make a transgenic mouse...in the lab, and have short generation times that make it easy to study the effects of genetic manipulations... be easily manipulated with tools like CRISPR to make transgenic lines. The fruit fly (Drosophila melanogaster...genetic material (Jennings, 2011).   The fruit fly makes a great model organism because they are easy and...C. elegans are also self-fertile hermaphrodites making it easy to conduct large experiments over multiple...60% of human genes have an ortholog in C. elegans making it ideal to study basic molecular biology processes...

... grow quickly and make more of the plasmids as they grow, scientists can easily make large amounts of ...guidance to understand the specific components that make up a plasmid and why each is important. Our “Plasmids...ability of plasmids to self-replicate within a cell make them attractive tools for the life scientist or ...particular environment Produce enzymes that will make specific, controlled changes to an organism’s genome...contains multiple protocols and troubleshooting tips to make plasmid usage as simple and straightforward as possible...

...and construction steps are complete, it’s time to make virus! The constructed viral vector containing the...particles are secreted into the cellular media, making viral collection fairly easy. Producer cell media...similar outcomes, so why and when should you opt to make a virus instead of just zapping your cells? That...transfect’ cells lines are amenable to transduction, making it a logical choice for efficiency. Where viruses... rarely done, viral pseudotype can be altered to make the virus specific for a type or subtype of cells...with alternative methods. Ready, set, transduce! Making virus can sound daunting, but it is easier than...

...arsenal to make sure people pay attention. Highlight your lowest points and highest points and make clear ...crucial part of interpersonal communication, which makes honing your ability to tell stories a similarly .... Whatever you’re working on, start wherever it makes sense to start and try to be as chronological as...protagonist In most cases, the protagonist will be you. Make sure to highlight what lessons you’ve learned and...failures. Foster conversation A story is not a lecture. Make yourself open to conversation — put your work on...

...find events stressful. Video chatting makes it pretty easy to make 30 minute appointments with people all...almost 6 months, you may be looking for more ways to make sure you are moving forward. Here are a few out-...hand at a video presentation? Most video platforms make it easy to record yourself. You can watch yourself...informational interviews, for more details on how to make the most of these exploratory conversations. If ...least two informational interviews a month you will make good progress on both building relationships and...

... are sensitive to blue light. Many have tried to make green molecules, but then alongside blue they actually.... So we developed a red molecule. We also had to make the blue light molecule much, much, much more light-sensitive... optogenetic tools in a deep part of the brain, making it sensitive to light. When they illuminated this... emotions, to see how a monkey changes decision making, to cure a human patient. We can't do it all by... I would say we still would like to find ways to make these tools more selective for different ions. Right...interesting. It would also be great if you could make neural silencers that require less power. Those ...

...techniques. Show up on time (don’t make your bench mentor wait on you!), make sure you are prepared (read any...success prior to your departure for the new placement. Make sure you read any relevant literature for the techniques...you have one) to communicate your plan as well. Making logistical and funding arrangements Visiting scientist... a structured visiting program, you will have to make arrangements on your own. This will include, but...a guest and likely a trainee in your new lab, so make a good impression! Remember that your host is going...

...can revisit the Q&A in the chat box. Presenting? Make it interactive! As a presenter, it can be difficult...giving a research talk, there are still other ways to make your session more engaging, for example, by using...to keep your video on and look into the camera to make it seem more like you’re connecting with the people...over. Depending on the conference, organizers may make the poster available after the conference is over...getting to know the app or virtual platform and making sure you stay connected. If possible, test out ...experience as an on-site conference, you really have to make space for it,” says Bachle. “On the flip side, you...

...important part of being a “good scientific citizen” and making sure you do what you can to help your fellow researchers...consistent way so other researchers can use them, to making sure that methods and protocols are available and... this communication and sharing fits together to make for a transparent and collaborative research environment...for being transparent in their communication or making their data and reagents readily available. What...confident that everyone’s enthusiasm for actually making it happen would increase dramatically. I’m not ...resources on the Addgene blog Find more ways to make your research more reproducible Read all of our ...

...over-diluting, and make sure to remeasure your titer after diluting to confirm. Luke Makana Hanley is a Research...Check out this post for a refresher on AAV titers Making contact with a cell Although an individual AAV ...see the dwindling proportion of AAV particles that make it through each part of the cell on their way to...additional best practices. Keep your cells happy - Make sure your cells are as healthy as can be to prepare...filter - When using a concentration filter column, make sure to rinse off the surface of the filter membrane...

... ability to make discoveries." If you ever run out of ready-to-use virus and want to make new lentiviral...genome editing tool is its developers’ willingness to make their CRISPR technologies available to the academic... reduced titer associated with large inserts can make lentivirus production frustrating - a researcher...protocol only to find that the low titer of her prep makes it unuseable. With the large size of the SpCas9 ...biology you're interested in studying. You need to make sure your assay is scalable and robust. What you...

...components. Making proteins outside of a cell - cell free extracts Typically, if you want to make a protein...more and more molecular biology-based activities making their way into high school classrooms, like gel...cost-effective, as each one is as cheap as 10 cents, making activities designed with FD-CF reactions affordable...they can touch. With BioBits, we want students to make the connection between the facts they’ve learned...experiences and knowledge with each other to help make learning biology more fun – bit by bit. Ally Huang...

...cargo is designed to make you sick…well, you feel it. But the same features that make these familiar viruses... and deposit a DNA/RNA cargo into their targets makes them attractive and tractable tools for genome engineering...essentially DNA (or RNA) delivery vectors, so what makes them better than the alternatives? Well, for one...Secondly, some viruses have a low immunogenicity which makes them ideal for applications with whole organisms... when introduced into an organism. This feature makes them an attractive tool for human subject delivery...

... aided design) drawings of their devices, which makes it very difficult to adapt the device to one's own...2020) or openUC2 (Diederich et. al., 2020) aim to make microscopy as a tool accessible to practically everyone...growing number of "community-driven" software that make everyday microscopy much easier. On platforms such...sources such as Amazon, Ebay, or Aliexpress, which makes it difficult to develop accessible open-source hardware...Distributing hardware via globally established makerspace networks and training users via workshops, seminars...newly developed techniques are the key enablers for making assemblies open-source. The sharing of digital ...

...we choose topics for the Addgene blog? Or why we make certain videos? In this post, we invite you behind...experiences from thousands of laboratories that makes up our Help Center, blog, website, social media,...process led to quick dives back into the literature to make sure absolutely everything was correct. Final touches...between our content team and our graphic designer to make sure things are clear, accurate, and in line with...shown.   However, for our blog posts, we usually make figures in BioRender or use appropriately licensed...review to catch those pesky typos, and hit publish. Making content is an iterative process. We are constantly...

...variable actually makes a big difference for the robustness of the findings.   How do we make research more... noise in the process of sampling and inference-making. Even perfect transparent and rigorous studies ...with a specific statistical test.  Preregistration makes clear the distinction between hypothesis testing...With Registered Reports, the authors are able to make that clear distinction between hypothesis testing...Furthermore, the journal and reviewers are incentivized to make sure that any possible null is really a true null...

... 2015. What makes lentiCRISPR v2 great? It produces a ~10 fold higher titer virus, making it ideal for...a point mutation in the puromycin cassette that makes it less efficient in some cell lines. pSpCas9(BB...V2.0 was made available through Addgene in 2015, making this the most recently available plasmid in the...a point mutation in the puromycin cassette that makes it less efficient in some cell lines so it may be... thank all of our depositors and requestors for making their reagents available and for supporting scientific...

...similar to previously discovered Cas13 enzymes - what makes these orthologs so special? The first property is...Cas13d enzymes have a median length of ~930aa - making them 17-26% smaller than other Cas13s and a whopping...whopping 33% smaller than Cas9! Their small size makes then easy to package in low-capacity vectors like...broader regulators of Cas13d activity. This property makes WYL proteins an intriguing counterpart to anti-CRISPR... yielded >80% knockdown. It seems that CasRx may make it possible to target essentially any RNA in a cell...and we at Addgene appreciate our depositors for making their tools available to the broader community....

...running one, you’ll know there’s a lot of choices to make when designing the experiment. What detection method...    Figure 1: The technical decisions to make when designing a western blot.  Psst! Looking...antibodies are relatively cheap and easy to source, making it easy to switch from one readout method to another...blocking buffer and a recommended concentration. That makes the choice easy! Protein-based blockers include ...Chemiluminescence kits When activating your enzyme, make sure to choose a chemilumescence kit appropriately...greater dynamic range than chemiluminescence enzymes, making it easier to accurately visualize proteins of wildly...proteins diffuse semiradially during the transfer, making membranes inherently “fuzzier” than gels. Quantification...

... unique cutting features and evolved PAM sites, making Cas9 an adaptable choice for many genome engineering... into crRNA). The size and simplicity of Cas12a makes it easier to package for therapeutic delivery by... to give it a go? The high specificity of Cas14 makes it a promising detection tool for rare, often hard...unlike Cas9, it targets RNA instead of DNA. This makes it particularly useful in generating transient edits...target sequence. The RNA-targeting ability of Cas13 makes it perfect for targeting mRNA and useful as a mechanism...targeting abilities of the other Cas family members make for useful implements in the scientist toolbox. ...

...light-gated ion channels such as channelrhodopsins (ChRs) makes them light-responsive. When light shines on excitatory...causes it to fire. Created with BioRender.com. To make channelrhodopsin-based optogenetics minimally-invasive...channelrhodopsins. Their new channelrhodopsins, called ChRgers, make the brain a little easier to access with optogenetics...injection. This lower multiplicity of infection (MOI) makes it challenging for ChR2(H134R) to function because...showed that ChRger2 stronger photocurrent properties make it better suited for systemic delivery with PHP....TL;DR The new channelrhodopsins, called ChRgers, make it easier and less invasive to use optogenetics ...

...your way. Label your tubes with a label maker. Label makers generate far more legible labels than your...into the lab wishing there was a machine that could make your life a hundred times easier. We’ve certainly...Addgene recently started automating a few protocols to make standard lab processes easier and more efficient...really liked that Beckman was flexible and willing to make removable parts to fit on the deck of the Biomek...pick up 8 tips and position the tips indepently making it easy to spread out your samples. This can be...

...mechanical work for editing. Class 1 CRISPR systems make up the majority of CRISPR-based immune systems in...broken down into different types, with Cas3 systems making up type I, the most abundant type in the Class ...away at both the non-target and target strands by making cuts every so often. Cutting both strands ensures... for Cas3’s biggest trick. Cas3 likes to bluff, making you into think it’s more powerful than it is. The...issues with expressing all Cascade components can make Cas9 easier to work with than Cas3. Future targets...1477. https://doi.org/10.1038/s41587-019-0310-0 Makarova, K. S., Wolf, Y. I., Alkhnbashi, O. S., Costa,...

... we’d like to make sure they are present at a very low concentration. So how do we make sure to catch ...an experiment because of poor reagent quality. To make sure our AAV vectors are of the highest quality,... mission of accelerating scientific research. To make sure that adeno-associated viral (AAV) vectors available...robots. Unfortunately, humans have been known to make mistakes. What if a tube is mislabeled or someone...K, Sanders E, Tasissa M, Kostman M, Tillgren M, Makana Hanley L, Mueller I, Mitsopoulos A, Fan M (2020...

...be “let’s try to make this STEM activity fun,” but rather “let’s see how we can make this fun activity...time is giving fewer examples and think of a way to make this section more interactive. If you can think ...good analogy that involves student participation, make it happen! How should we use CRISPR? How shouldn...students that scientific history was very much in the making. One of the exciting, and challenging, aspects ... Perhaps you have young children and you want to make sure their STEM education is supplemented. A 2009...

...reliable and robust high quality viral vector service, making an array of ready-to-use viral vectors available... prioritization process to decide which tools we make available. This prioritization happens for good...of our most popular ready-to-use viral vectors, making up almost 10% of our requests by 2019 (Figure 5A...which preps to include in our service allows us to make ready-to-use viral preps that are available quickly...Hub before selecting an AAV can help a researcher make a more informed choice on which AAV to request and...since, we’ve learned so much about what it takes to make these viral vectors not only available, but accessible...

...to be a part of projects that connect people and make you work in a collaborative environment. Think of...conversations about your ideas and don’t be afraid to make something concrete happen. You may have more willing... institutions personally on conference calls and make sure to maintain these relationships by keeping ...organization. You can meet collaborators  for lunch, make a point of talking to them at a seminars, or even...from an internal to an external connection is to make sure you keep open lines of communication with colleagues...see interesting events, articles and newsletters I make sure to share them with a short note - this creates...

...Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, Essletzbichler P, Volz SE, Joung J,...Non-SpCas9's bind a variety of PAM sequences, which makes them useful when no suitable SpCas9 PAM sequence...non-SpCas9's may have other characteristics that make them more useful than SpCas9. For example, Cas9 .... BV3L6 Cas12a, AsCpf1, uses a 5'-TTN PAM which makes it easier to target AT-rich genomes (Kleinstiver...Gootenberg JS, Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F (2015) In vivo...

... Denotes the presence of an additional plasmid-- make sure to maintain this by growing on appropriate ...Induction timing is important, as you typically want to make sure your cells have first reached an appropriate...exponential growth phase are alive and healthy, which makes them ideal for protein expression. If you wait too... as there are not enough cells in the culture to make protein.  The DE3 lysogen/T7 promoter combination...positive selection and a p15A origin of replication, making it incompatible with other p15A plasmids. pLys ...Double-check your plasmid backbone and expression strain to make sure they are compatible. An arabinose-inducible...

... can promise is that my suggestions can help you make the best case for your candidacy. A major reason...complains that a seminar was too easy to understand. Make your seminar accessible. Let the experts ask you...minutes of the chalk talk. Realize that it is rare to make it through the details of all of the aims of a chalk...points you want to convey for each section or aim. Make sure those get priority. The fine details for aficionados...examples are not especially helpful. I set out to make something practical and accessible. If there's something...postdoc salaries all too well. If there's a group to make money from, postdocs are not that group. If you ...

...even one less run a week makes a big difference. Clean out freezers so you can make room and don’t have to..., check out my Recycling and Innovations posts. Making small changes in your daily habits can feel ineffective...sustainability problem, how can one little change make a difference? Individual changes to our behavior...wouldn’t leave your oven on all day to save time making dinner at night, so why leave your lab equipment...

...requires that the RNA be tagged. “This tool makes it possible to make the protein specific to any endogenous...PUF proteins! A newly available PUF Assembly Kit makes it possible to devise RNA binding proteins to hit...continue to evolve and improve, Abil says they will make those new plasmids available through Addgene, and...the final version,” she said. “If we learn how to make the library even better, we would be very happy ...

... puncta in each cell. When making figures for a paper or presentation, make sure the images are accessible...want to look for them? Or maybe you just want to make some really beautiful images? Multiplex IF may be...different colors in one session – fast and easy! Just make sure your primary antibodies are all of different...considerations to keep in mind. Most importantly, make sure you are using primary antibodies from different... and dye meets your needs and budget, or you can make your own. For example, to label the rat brain sample...