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...three or four guides directed towards that gene as there can be a large difference in guide potency. Thus...whichever guides are closest to the transcription start site (TSS). We recommend targeting the guides to a ...scientific world by storm. While Cas9 has been most widely used to create specific edits in DNA, there has...required for Cas9-based activation. CRISPRa for genome-wide studies One of the best uses for Cas9 activators...  Which Cas9 activator should I use? There are a wide variety of activators you can use for your experiments...transcription. That being said, we generally pick guides by putting the promoter of the gene into a gRNA...(2015) WU-CRISPR: characteristics of functional guide RNAs for the CRISPR/Cas9 system. Genome Biol 16:...

...dumpsters outside the open windows in the middle of your talk. Technical difficulties turn your slide deck ...interest incorrectly on slides 8-16. Your notes disappear. The wrong version of your slide deck gets uploaded...hard. Aside from the diving board charades, I doubt anyone but me remembers any of the incidents above–and...Addgene blog Transferable Skills Guide: Time Management Transferable Skills Guide: Conflict Resolution Transferable...stand up in front of an audience and convey your ideas clearly and strategically is powerful in almost ... allows you to share and build support for your ideas, invite others into decision making processes, and...help me achieve my purpose? Should I go over this slide again? Say, 'Oh, I forgot to mention X, so let's...

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

...investigator who is using an in vivo animal model should consider the possible elicited immune response from viral...AAV. While these benefits make AAVs useful for a wide variety of experiments, care still needs to be taken...particular AAVs to use for your research. Below I’ll provide some tips that will help you think about how to...from https://www.addgene.org/viral-vectors/aav/aav-guide. Where in the tissue/what cell type? In the CNS,...distance traveled, etc.) (4). This has also been identified with AAV2 (5), AAV6 (6) and AAV5 at excitatory... many experiments, a group of researchers has identified this feature of AAVs as a new tool for neuroanatomical...miRNAs), short hairpin RNAs (shRNAs), antisense oligonucleotides (ASOs), zinc finger nucleases, and many others...

...vectors Although retroviruses are still in use, considerable efforts have been devoted to develop efficient... Inducible lentiviral transfer vectors The most widely used inducible lentiviral vector system is the ...manipulated with care. See the NIH’s biosafety guidelines for more information and always be sure to consult...another. For experimental consistency it’s a good idea to purify and titer your virus. Virus purification.... Don’t hesitate to check our lentiviral vector guide to get more information and to get access to additional...Order Ready-to-Use Lentivirus Browse Our Lentivirus Guide Pages  ...

...Today in our transferable skills guide: Collaborating with others outside your organization also known as...communicate outside my scientific expertise. Organizing a career fair for early-career scientists provided a crash...Interestingly, the ability to work with others outside one’s organization was found in the aforementioned...important in a non-research context, especially when considering small to mid-sized companies and organizations... Addgene as a nonprofit organization is very considerate and thoughtful about its limited resources. Rather... curve for collaborating and it is always a good idea to start close to your comfort zone and slowly expand...Collaboration opportunities within an organization but outside your comfort zone As part of my position at Addgene...

...comprehensive guide to Conflict Resolution. There are countless classes and resources that can provide details...ScientistsTransferable Skills Guide: LeadershipTransferable Skills Guide: Teamwork  ...” in quotations here because I would not have considered myself neutral in any of their disagreements....open new career opportunities for you, if you can identify them in yourself and learn how to sell them. Types...decision making, and stalled projects. You want to identify and address these types of harmful conflict early...other parties, the first step I always take is to identify the real source of the conflict. It can be easy...important to retain the stories and examples that have guided your resolution processes in the past. Types of...

...what I had done in order to identify potential sources of error and then decide on a strategy to fix the ...from the transferable skills guide: Teamwork Collaborating with others outside of your organization Time...need some help. Today in our transferable skills guide: Problem-Solving. I don’t know about your experience...went wrong with a PCR is a pretty niche skill; outside of a lab, where is that going to come in handy?...has been invaluable in my more recent positions outside of the lab. Granted, I currently work on the Scientific...situation, come up with potential solutions, and decide on a course of action that effectively resolves...solving as a generalizable process.  Step One: Identify the problem and gather information to help you...

...college, I knew I wanted to try research, but had no idea what to expect. I had a lot of help from my peers...scientific endeavors. I hope that this post can provide some perspective and advice to any undergraduate...no biological or biomedical scientists and had no idea what the daily life of a scientist was like. The...importance of research in basic biology, medicine, epidemiology, public health, and more.     A few of the...professional training after earning a doctorate Considered a "stepping stone" position to become a professor...future. How do I get started with research? I had no idea what my scientific interests were or what I wanted...is good advice, and it is ultimately up to us to decide whether we want to take it or not. Choose wisely...

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

... The Addgene Videos Page 5 Science Rap Videos and Their Creator: Tom McFadden Using Video to Share Your... we’ve gained along the way to ourselves so we decided to share some of our tips in our new series: Addgene...Have some suggestions for future tips?  Send your ideas to [email protected].  Additional Resources on the...Additional Resources on Addgene.org Browse our Video Library Check out Our Protocol Pages Find Molecular...

... that one on one discussion can be confidential. Finally, provide timely feedback. We often associate...need some help. Today in our transferable skills guide: managing a team. You’ve just been promoted, congratulations...advance as much as possible so I could more easily identify what to delegate and to whom (make sure that person...expectations, answer any questions, and (most importantly) decide on a timeline. My experience is that tasks often...of X’s work for a couple of weeks now”. I had no idea this was happening even though I met with X on a...when having a mentor - someone more experienced to guide you - is valuable. Protect your team Make you team...impact your work and your life. It’s up to you to decide which one you want to be!  References Becoming...

...need some help. Today in our transferable skills guide: Time Mangement for Scientists. Back when I used...my own experiments in graduate school, I had to divide my time between lab work and practicing my SciComm... but if you ever find yourself in need of a few ideas, I hope you’ll find this post useful. After a bit...which tasks to actually do, you should err on the side of things that have the most utility; i.e. those...according to my priorities, I have a pretty good idea about the specific things I’ll be doing through ... it’s enticements are far too distracting. (As a side note, I’d recommend the book, “Bored and Brilliant...

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

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

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

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

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

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

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