...run in triplicate.  The nice thing about qPCR is that since it measures relative quantification - that... not need to be run in triplicates. ddPCR is an extremely accurate application, even in areas of the genome...Digital PCR for AAV QuantificationPolymerase Chain Reaction Overview and Applications Resources on Addgene.org...blog post, we're going to focus on PCR-based applications (qPCR and ddPCR.) qPCR Quantitative/real-time...range of RQ values as noted above.   In both applications, results can be affected by DNA quality and ...absolute quantitative data, instead of relative quantification.  Even though no one option is perfect, with...your variant options, you’re likely to find an application that matches your needs. Happy quantifying!  ...

...after credible attempts to replicate major findings could not be replicated by other researchers.There...published research is difficult to replicate.  Why is replicating previous research findings so difficult...can be difficult to replicate. Unclear methods Until someone attempts to replicate the results of another...positives in original studies and false negatives in replications are unavoidable, but can be minimized with high...obtaining a different result after conducting a replication study. An important difference between the original... methods, QRPs, and small samples, then when replications do not come out the same as an original research...data, materials, and code Only when you try to replicate the work of other researchers (or your own from...

.... I needed to a) figure out the sources of miscommunication and b) find practical solutions.  My PhD ... for this task. Good scientists learn how to communicate their work differently based on their audience... family is going to vary widely. The way you communicate your science at a poster is going to vary depending...two main problems with Addgene’s cross team communication. Although these real world examples did not ... for the container to be used in cross-team communication A list of “AKA” names, or jargon terms, for ...tracking system. Though you might not have to communicate with a team of software engineers in your lab...experimental protocols. Clarify: Asking for clarification during a meeting can help get everyone on the...

...and applications.    Plant applications We outlined many synthetic biology MoClo applications, but...- and multi-gene constructs for a variety of applications. MoClo tools can be used in bacterial and cell-free...and the kits Addgene offers to kickstart these applications, no matter what your model organism is. MoClo...this blog, we will highlight some common MoClo applications and kits. Keep in mind that this technology ... it, MoClo might help you achieve it! MoClo applications and specialized tools Synthetic biology in bacteria...and multi-TU constructs for synthetic biology applications in bacteria or cell-free systems. The approach...combinations of promoters, terminators, solubility and purification tags, connectors, cDNAs, and more to find the...

...genetic underpinnings of these processes has been complicated by the lack of precise genetic tools for modulating...recent advancements and offer commentary on their application to neuroscience research. What is CRISPR-based...accessible to neuroscientists, not all neuroscience applications require such optimized tools. This is elegantly...necessitates such molecular gymnastics for sensitive applications such as CRISPR screening. The plasmids used ...used by the Qi Lab for this particular application can be found here.  Why CRISPRa? Although cDNA overexpression...gene activation in select in vitro and in vivo applications, the tools described above make CRISPRa in the...relatively easy and accessible. There are also other applications for which CRISPRa is well-suited, including ...

...some tips for performing this purification.  Check out the AAV Purification protocol page for more detailed...! Subscribe to the Addgene YouTube page! AAV purification by iodixanol gradient ultracentrifugation This...This video goes over AAV vector purification using an iodixanol column gradient. We'll show the loading...

...Further, when science is communicated in a format that is accessible, that communication often neglects the ...SciCommSummer Scicomm Series: Modes of CommunicationCareers in Science Communication: Science Writing... to save a few days’ worth of capsules to self-medicate with when they get a cold. A primary school teacher...dihydrogen monoxide (H2O). Studying science communication as a PhD student Fortunately, by the time I ...Otago had established the Centre for Science communication and I was accepted as a grad student. I chose...a phenomenon called “motivated reasoning,” the catchall term for a range of neat little tricks our minds...found opportunities for more hands-on science communication. Between New Zealand’s multiple Covid-19 lockdowns...

...Riglar Advocating for Science Symposium and Workshop 2016 – Sept 16-17 MIT, Boston The Advocating for Science...futureofresearch.org/advocating-for-science-boston-2016/. Sign Up For the Advocating for Science Symposium...Director of Media Communications, Harvard University Ray Howell -Founder of Howell Communications Philip Brenner...union dues. Time will tell the extent of the implications of this ruling and whether graduate students...labor laws, rather than as trainees in a special category, as has been the ‘labor-of-love’ philosophy of...tight lab budgets and even simply being able to categorize all postdocs at institutions, will be challenging...Revisited” 2014 Appendix B p93-95). For those advocating in this area, keeping the pressure on for further...

...DE3. For Phagocata morgani and Phagocata gracillis, the process is a little more complicated. These strains...Girardia sp., Dugesia dorotocephala, Phagocata morgani, and Phagocata gracillis. There are 11 different ... coli. Thus, to get the dsRNA into Phagocata morgani and Phagocata gracillis, one has to purify the dsRNA... in situ hybridization experiments. A recent publication [1] from the Sánchez lab describes the plasmids...the deposit) are described in the Sánchez lab publication [1] so that the students performing these experiments...mRNAs are subsequently destroyed by Argonaute-catalyzed nuclease activity or their translation disrupted...-amputation planarians after knockdown of beta-catenin. Other gene targets do not require amputation to...

...science communication are as varied as the field itself. Among other things, science communication comprises...certain aspects of art and education. If you suspect that science communication might be for you, don’t ... science education Do you aim to reach young people? Try networking with your local education community... Training – Get a formal education Some people get into science communication by taking a media or journalism...project. Science communication opens doors Getting involved in science communication has been the most...career researcher looking to break into science communication? Everybody goes about this in their own way....t wait. Start communicating now. These 9 strategies will get you started:  Subscribe to Career Blog Posts...

... Working on the Cat Microbiome Project Another project we are working with is the cat microbiome project...data for analysis. And, well, cats... everyone knows the internet loves cats! This is mostly a project to...gut of the domesticated housecat, and seeing how that makeup differs from that of wild cats with the goal... goal of optimizing the health of domesticated cats and learning about feline microbiomes. One of the ...OpenWorm Project Website: http://www.openworm.org/ Cat Microbiome Project Website: http://www.kittybiome.com...biologists and computer scientists learning and communicating with one another.      Currently our main focus... of Minnesota has declared our makerspace an educational institution meaning that we can obtain known ...

...where readers may locate data supporting the research reported in PLOS publications. The policy was not... is founded on the principle that scientific publications must contain enough information (or links to...probable to some extent - but that is the price of publication. Who is to say that the initial data-generating...logically annotated and arranged, as in an actual publication and not require interaction with the data authors...likely, at least in part, due to our inadequate cataloguing of how results were obtained and this alone is... Nature Publishing Group recently launched a publication called Scientific Data, which aims to act as ...how they view the information. We are used to publications presenting a polished, standard and usually ...

...they asked me, 'How do you feel about dissecting a cat?' And I’m like, 'Well, I’m here so let’s do it!'"...in creative science education tools? Check out this Addgene podcast with educator and science rap mastermind...In this post of the Careers in Science Communication blog series, you’ll hear more about Caitlin Runne-Janczy...subject matter expert (SME) at eScience Labs, an educational company that creates hands-on science lab kits...jack-of-all-trades. “As a subject matter expert in the educational sphere, it can range from researching an area...take a step back and look at new technologies in education and ask ‘Can we implement that?’ [And think about...business.” Caitlin’s advice for budding science communicators: “My recommendation would be to take any opportunity...

...In this post in the Careers in Science Communication blog series, you’ll learn about Caitlin Runne-Janczy...Product Development Manager at eScience Labs, an educational company that creates hands-on science lab kits...through Caitlin’s blood. “I come from a family of educators, mostly elementary school, middle school, and ...where I came from.” Working at eScience Labs, an educational company that creates hands-on science lab kits...when it comes to the labs.” While her love of educational science writing is clear, Caitlin’s career plans...to do scientific editing.” It was for Cactus Communications, a company that provides scientific editing...can learn what it’s like to work for a science education company. And make sure to subscribe to the Career...

...about careers in science communication. When I started my Science Communication Internship with Addgene...ll interview three science communicators who work in the biotech, education, or nonprofit industries. ...re interested in learning more about science communication careers, you’re in the right place. For this...should be. He debated taking a position as an application support/sales representative in industry or a...job. “I am really more of a multimedia science communicator than a science writer these days. I do some ...illustrations for summarizing mechanisms and applications that are relevant to things IDT does. I also...next couple of months. This involves a lot of communication with the presenters, and editing of abstracts...

...see any signs of DNA in your final tube after purification. How do you know if you actually have DNA in...you choose could be based on your downstream application, time, and instrument availability. UV absorbance... at 230 nm. The A260/A280 ratio is used as an indicator of DNA purity. Ideally, this number should be ...like a PCR product). This method also gives an indicator of DNA or RNA contamination based on the presence...before larger fragments. Figure 1: The quantification of a RNA ladder using capillary electrophoresis... about quantifying DNA In choosing your DNA quantification method there’s many things to consider: cost...

...things or communications planning.” To beef up her resume, she decided to complete a certificate in Strategic...Podcast Episode on Science Communication Check out Other Science Communication Posts Resources on Addgene.org...In this post of the Careers in Science Communication blog series, you’ll hear from Susan Keown, a staff...Cancer Research Center. Susan Keown’s science communication journey started with something unexpected: termites... time position with a Washington D.C.-based communications consulting firm, Palladian Partners. “I was...Strategic Communications and Public Relations at the University of Washington and the connections she made...storytelling is an impactful tool for science communicators. “The power of a story is to make a human connection...

...scientists have long used antibody based affinity purification and mass spectrometry approaches. Recently however...that are tagged with biotin using avidin coated purification techniques. Due to the high affinity of biotin...challenge for classic antibody based affinity purification/mass spectrometry methods.  BioID: a promiscuous...can even target BirA* to specific subcellular locations, such as the nuclear envelope, allowing scientists...scientists to probe interactions in locations of interest.  Try BioID for your proximity labeling experiment... proximity labeling (Rhee et al., 2013). APEX catalyzes the oxidation of biotin-phenol to the short-lived...mutation (R40G) in a conserved residue in the biotin catalytic domain to allow for promiscuous biotinylation....

...#1: Affinity tag purification You tag YFP and use an affinity column for purification. After binding YFP...  Light controlled protein purification with OptoNBs The first application for OptoNBs takes us in vitro...light-controlled affinity purification of unmodified proteins. Affinity purification methods typically require...It’s time to choose your own protein purification adventure. You want to purify your favorite protein...elute YFP.  Option #2: Opto-Nanobodies (OptoNBs) purification You skip adding a tag to YFP and instead use...sound like fiction, but light-controlled protein purification is now a reality thanks to OptoNBs. Recently...response to light allows for tagless protein purification as well as reversibly regulation of endogenous...

...phosphatase-accessible. Kinase Translocation Reporters (KTRs) Kinase Translocation Reporters (KTRs) are genetically...provides an alternative tool with their Kinase Translocation Reporters (KTRs) whose cellular localization...activity is enhanced, leading to cytoplasmic translocation of the reporter. As a result, the KTR is shuttled...nuclear export and import rates can influence KTR translocation. Work from the Covert lab suggests that these... negligible effect on KTRs, however nuclear translocation rates are another variable to consider when ...measurements of kinase activity in single cells using translocation reporters. Nature protocols, 13 1, 155-169. ...