...analogous to traditional restriction enzyme cloning. Sticky-end mediated gene transfer would be particularly...indicated below each graph. Figure adapted from Kleinstiver et al., 2016. Until recently, two major elements...off-target editing efficiency. Kim et al. and Kleinstiver et al. characterized genome-wide editing efficiency... to single mismatches, but variably so, with Kleinstiver et al. reporting that Cpf1 can tolerate mismatches...essential function in Cpf1-mediated editing, as Kleinstiver et al. saw no decrease in Cpf1 activity with ... off-target editing rates, determined using sophisticated genome-wide analysis. At many of its computationally...activity than that previously observed with SpCas9. Kleinstiver et al. suggest that AsCpf1’s off-target rate ...

... that cannot be distinguished using conventional flow can (sometimes) be distinguished using spectral ...applied to “subtract” this spillover and correctly distinguish between markers. Additionally, since the emission...hand, utilizes the full spectrum of light to distinguish one fluorophore from another. Instead of using...algorithm and single stained reference controls to distinguish one fluorophore from another within a fully stained...These unique peaks in the spectra can be used to distinguish these two fluorophores from one another during...of “0” indicating those markers are completely distinct from one another, and a value of “1” indicating...those markers are identical and completely indistinguishable. This tool is of particular value when designing...

...Resources National Cancer Institute: Cancer Statistics Page National Cancer Institute: Ras Pathway v2.0 Page...cancer-related complications each year. Despite these grim statistics, new strategies for attacking cancer are yielding...undergo programmed cell death while simultaneously stimulating growth and division. The combined effect is unregulated...so, to pose the greatest threat, cancerous cells still need to successfully infiltrate the circulatory ...as those within hair follicles and lining the digestive system, causing discomfort and illness. Further...involved in the Ras pathway alone and thousands of investigators studying them, how do we ensure that the research...Cancer Research Plasmids and Resources page, investigators may select a pathway and identify any genes ...

...programming life. Which pieces are still missing? Endy: That's a great question. You can see that we have the...substantial. It's estimated that genetic engineering as applied to products on the domestic market generates...programming languages like Java or Python. The question is who owns languages? Who owns English? Well,...every year. That's two percent of the U.S. gross domestic product – equal to mining. There is an absolutely...The parts we are releasing simply plug into this existing economic activity. People are using BioBrick parts...I'd leave it is mostly to the imagination. The question is: what can you use biology to make? If you think...-gene system to do something. We know from our testing of these parts that the chance your genes will ...

...at the Broad Institute of MIT and Harvard, Dana-Farber Cancer Institute, and Koch Institute, where an exploration...any other ancestry group. With the existing methodology, they estimate that the African ancestry-associated...and well-documented history of racial bias, which still impacts and influences current research. Understanding... Aaron knew that their study had significant statistical limitations, and expected to find only a few,...Perturbation Platform to design a new library that is agnostic to ancestry which will reduce the likelihood of...five-team response to address this issue, but it still points to a far larger and more complex problem ...Paul S. Appelbaum, Bege Dauda, Anna Di Rienzo, Agustin Fuentes, Stephanie M. Fullerton et al. Getting ...

...particular scientific question - even when others have already answered this question and used it to publish...optimize our vector - overall, I spent two years testing and reformulating our AAV construct to optimize... of that data was published.    Fig. 1: Testing of three different AAV serotypes (AAV1, 8, 9) injected...scientific support team answers hundreds of AAV questions every year on a range of issues related to AAV...many from people trying to answer the exact same question I was: What AAV vector is best for my specific..., we must therefore actively work to remove the stigma around sharing negative data and instead focus ...Recombinant AAV Produce Chemically and Functionally Distinct Vectors. Molecular Therapy - Methods and Clinical...

...Addgenies. Got a question at 9:00am GMT? We’ll be poised and ready to answer your questions about Addgene...counterparts are still sleeping). Plasmid sharing in Europe: Addgene’s request statistics Since 2004, Addgene...Europe. Despite these fantastic numbers, a lot of European scientists and institutions are not aware of what...Addgene has been there to answer your plasmid questions. But until now, you’ve always had to call the ...75,000 plasmids to more than 1,500 different institutions in Europe. In the last four years alone, the...Years of Plasmid Sharing: Interested in more statistics from Addgene's 10 years of plasmid sharing? ...

...the first time, have lots of questions. To keep up with all these questions, Dr. Le Cong and Dr. Feng ...most popular questions and answers. Here are just 4 of the CRISPR-Cas9 FAQs. If you still have more questions...CRISPR technologies and their applications can be exhausting. A quick search for "CRISPR", short-hand for ...developed a Google Forum to allow scientists to ask questions and get answers on the fly. But just like the ...expression, though it is possible for the plasmid to still express without the G. Because the G is only one...CRISPR protocols, and gRNA design tools Need more questions answered about CRISPRs? Check out the full list...

... focus on how to choose an advisor/principal investigator (PI) that will also serve as a good mentor. ...advisor is a good mentor? You have to ask a lot of questions. Personality and style conflicts are a common ... determining a good fit. Here are just a few suggestions for determining whether an advisor will be a ...potential advisor and don’t be shy about asking hard questions. Talk to as many lab members as possible. Try ...Ask alumni of the lab. Most labs have websites listing alumi and it is easy to use publication records...this blog post. This document includes a list of questions to ask your potential advisors and other related...Check Out Angela Depace's Talk on "Precision and Plasticity in Mentoring" Don’t ignore warning signs If lab...

..., there still remains the problem of prevention. Asbestos exists pretty much everywhere, still, in old...genetic factors may be in play. However, there is no question that asbestos exposure remains the single-most...Asbestos, and certain other naturally occurring asbestiform minerals such as erionite, are the only scientifically...scientifically proven causes of the disease. There is no question on this point within the medical and scientific...response. This important biomarker can be used to distinguish between three groups of people:  Those who have...different approach to immunotherapy is bevacizumab (Avastin). This drug (known as an angiogenesis inhibitor... centers in the world. The development of new diagnostic methods and treatments such as these has brought...

...manipulation still need to be developed for each organism, but researchers are optimistic that this can...recently was invited to attend the first Principal Investigator meeting of the NSF EDGE (Enabling Discovery ...food. They have chemical pathways generating interesting drug-like compounds especially cardenolides which...traits with good genetic mapping morphology and interesting behaviors. There is diversity in the social bonds...to get edited embryos (replacing the nucleus is still very difficult). Cuscuta (James Westwood) Dodders...from other plants and are unusual enough to be interesting biologically. They have economic importance as...Gasterosteus aculeates (Dan Bolnick) A paper on stickleback fish science was first published in 1867. It ...

...lead to under and overestimation of fusion protein activity. We encourage investigators to characterize ...Erik L. Snapp and Lindsey M. Costantini. "You underestimate the power of the Dark Side." --Darth Vader in...While Vader was referring to the evil side of a mystical "Force," this quote is equally applicable to many...the cytoplasm. That is, unfortunately, some investigators realize too late that they have missed the impact...behaving properly, FPs enable researchers to investigate the localization and dynamics of fluorescent ...environment where it will localize [4]. Most investigators are surprised to learn that FPs are often poorly...most eukaryotic) proteins localize to chemically distinct subcellular environments, including the organelles...

...Woodgett. Public Library of Science (PLOS) created a stir earlier this year when it announced its data access...caused a storm as the organization left open the question of how much data was necessary and reasonable.... errors in assumptions, use of inappropriate statistical methods, omission of some data, etc.  The firestorm... lab has efficiently trawled their own data? A distinct (perhaps similarly impoverished lab) might apply...published, this data is usually condensed into a statistically evaluated set of values.  However, to evaluate...what was actually carried out. While no one is suggesting all data be stored in a retrievable format, we... images are said to be representative or the statistical variation is calculated and placed in the legend...

...overcome. Conflict, and its inherent tension, is interesting. It’s what drives a story forward - and this ... that the superheroes win in the end. But we’re still invested in seeing how they do it.    Finding...your formal scientific communications, they are fantastic fodder for science communication. The trick is... the pieces that make you think, “Oh, that’s interesting! How did you solve that problem?” Those are the...big-picture problem, by asking your audience a simple question, like “Have you ever thought about how X works...conflict, you’re encouraging your audience to ask questions. Why didn’t the experiment work? What did those...preventing them from moving forward? Answering these questions will bring you and your audience - via the scientific...

...What tools are still missing from the optogenetics toolbox? Boyden: That's a good question. We've made a...about this latest optogenetics advance, the big questions that motivate his interest in neuroengineering...neuroengineering, and some of the optogenetic tools that are still missing. Optogenetic tools for neuroengineering Addgene...compete against each other. Getting two-color stimulation has been a holy grail of the field. The problem...fundamental understanding of the brain, which has humanistic and philosophical implications. As an example... a lot of progress. I would say we still would like to find ways to make these tools more selective for...calcium or some other specific ion that would be interesting. It would also be great if you could make neural...

...Harvard Business Review article on procrastination recommends testing out how much time you can possibly...conceptual chunks: prioritization, planning, and procrastination. Time management through prioritiziation Every...option. Even if the full experiment fails, you’ll still have made small accomplishments each day that should...Eric before doing anything else. Preventing procrastination Even if you’ve done all the prioritizing and...and planning in the world, procrastination can easily prevent you from making progress on a task. For ...few things I’ve found useful to avoid such procrastination: Remove distractions: Probably the easiest...book, “Bored and Brilliant” for learning more fantastic reasons to remove yourself from distractions every...

...et al. "The intestinal stem cell markers Bmi1 and Lgr5 identify two functionally distinct populations....Stochastic multicolor labeling is a popular technique in neuroscience and developmental biology. This... persistently fluorescent during development. Stochastic multicolor labeling systems, many based on Brainbow...for improving the level of detail possible for stochastic multicolor labeling of the brain. Labeling Drosophila... Cre recombinase can be used alongside common, existing tools for fly research that use Flp recombination... to describe the self-renewal of cells in the intestinal crypt, and since then has become a powerful tool... researchers who developed Tetbow. Conclusion Stochastic multicolor cell labeling systems allow single...

...being able to ask follow up questions, or ask people to rephrase questions was key. I had also hoped that...that’s what keeps things interesting. And hopefully they’ll be just as interesting as the finished product...familiar with this, it’s a method for building and testing new products or methods, using metrics and feedback...webcams.  Build: I sent the participants interview questions they could answer, and I’d cut together a video...the office had partially reopened. But there were still many restrictions in place, most importantly, masks...would allow me to set up a camera in a room, but still have control over focus, zoom, stop/start record... to mount a battery to it if necessary. I could still control the camera from a distance if I needed to...

...listeners to ask questions and record those that were asked. Reflect on the questions and come up with...Google.slides or Powerpoint file right now to capture interesting key results, striking images, creative ideas,... write itself when the time comes. 5. Choose interesting content Talk about something you know a lot about...commercialized, just talking about the possibility is interesting. Check out Joanne's Reddit AMA 6. Practice, practice...everybody loves a great animation. 8. Predict questions, prepare answers Speaking pros always have a few...always better than too long, and ensures time for questions. Don’t be tempted to cover too many things. One...this enough…more is almost never better. 10. Be enthusiastic You don’t have to emote beyond your comfort ...

... to the products of parental plasmid digestion (blue). Digestion with enzyme “B” detects primer multimerization.... Many thanks to our Guest Blogger Kristian Laursen! Kristian Laursen is a researcher at Cornell University...This post was contributed by guest blogger, Kristian Laursen from Cornell University. Site directed mutagenesis...can be used to introduce specific nucleotide substitutions (or deletions) in a tailored manner. The approach...perform alanine scanning mutagenesis or targeted substitution of key residues), and in SNP analysis (to introduce..., the template is removed by DpnI restriction digestion leaving behind only the mutagenized plasmid. Although... bases at the 3’-end interfere with plasmid reconstitution, and consequently Taq polymerases cannot be...