...you a science student or early career researcher looking to break into science communication? Everybody ... festivals and outreach If you are comfortable talking in public, volunteer at science festivals. Or engage...education Do you aim to reach young people? Try networking with your local education community. Get involved...environmental groups, charities and NGOs are often looking for outreach volunteers as well. 4) Blog - Hone...’t you record a podcast or put some effort into making a video? Use your unique talent to communicate ...Association allow you to spend two to six weeks working at media outlets such as the Guardian, BBC Breakfast...education Some people get into science communication by taking a media or journalism course. Courses can help ...

...programs is almost as daunting as the program itself. Picking a mentor comes with many challenges, and the flood...nosy question! The Application Process Broadly speaking, the PhD application process tends to go something... to potential mentors?” “Where do I start with picking universities?” and “How does COVID-19 impact all...program is the best fit for you and your goals. Making a List of Schools Creating a list of schools is...chance to get to know the faculty that you might be working with for the duration of your program and ask any...information about the campus even if you can’t visit. By talking with people at the institution, you can get a better...into what matters to you. It can also be nerve wracking to ask questions in front of other prospective...

... projects and they end up making 30 constructs. They’ve never done any kind of cloning and they don’t ...tools you are working on for plants? Patron: Our lab and others at the Sainsbury are working on a lot of...general. We are funded by a charity and since we are making all the tools for plant research here and validating...problem. MoClo is really a modern toolkit for those working in plants, including many things people already...Addgene: Do you have any particular tips for those looking to get started with MoClo? Patron: The parts will... that you need from the kit and getting started making your own constructs with Golden Gate MoClo. The...network here. She talked about the advantages of talking to people and representing yourself as a real person...

...’ve gained experience working with similar protocols, and look - they’re seeking someone who has, to a... a specific field. While searching for jobs and making connections, avoid falling into an identity trap...retrieved your lucky socks, persevered through a nerve wracking commute complete with a disabled train, and now...this job was made for you. They must have been looking at your resume while they wrote the job description...side of an issue. Maybe you also showed critical thinking skills by dissecting their problem and piecing...organizational skills. If you are responsible for working with lots of students, managing inventories in ...pretty much any other workplace. To get your mind working, take a look at the short list of transferrable...

...because you are not (yet) asking for a job or a big favor - you are generally asking for some advice and guidance...researchers do learn MANY transferable skills while working in the lab and by offering advice on areas where...many methods and machines one team can learn so linking together expert teams can push a scientific project...graduate studies at Karolinska Institutet I started working with other groups. First within my department where...meet collaborators  for lunch, make a point of talking to them at a seminars, or even set up regular meetings...much more effective than simply sending a message asking how things are going. Science careers where the...studies in regard to reaching out to new people, checking in regularly, and maintaining useful notes that...

...authors’ compared nicking the top strand first followed by the bottom strand versus nicking the bottom strand...position. The “sloppy” PCR reaction is created by making some or all of the following modifications: 1) ...packaged into phage and then extracted. Besides taking a couple days to generate the phage-derived uracil...ready to be tested in a functional screen of your liking. This method is similar to site-directed mutagenesis...with a twist since it relies on the sequential nicking, degradation, and PCR-generation of a mutant version...Supplementary Note 1 in Wrenbeck et al for details on how nicking was optimized. How should I design my mutagenic...gene of interest has a BbvCI site(s). If so, the nicking mutagenesis protocol will still work if the orientation...

...step by step instructions on creating a budget, making decisions based on that budget, and negotiating...timeline Start with your thesis in mind. Imagine the kind of data your dream discovery will need, and how ...should reflect salaries in the region you’ll be working. At this point, your budget bottom line costs may...position. This is important to ensure that you’re looking at institutes that have the means to support your...values. All too often, we see early investigators making unnecessary short term sacrifices that hurt their...dependent on a pathology microtome/cryostat, then making a careful decision on the condition and availability... and limitations. Everything is negotiable. Try asking for discounts, collaborations, access to internal...

...sgRNA design is not possible without relying on some kind of pre-analysis. At the end of 2014, I began developing...contents spill to the ground. There is something akin to a frenzy as the crowd tries to weigh and determine...each item changes from what the reviewers so painstakingly assessed to something a bit more representative...community as a whole has yet to settle on any true kind of value. I’ve dubbed this the “Piñata Effect”, ...question is then, should we use all tools as some kind of a meta/Frankenstein app? Or perhaps there is ...2013 about 11 tools have been added every year.   Breaking down the barriers: My view on current sgRNA design...released with advanced functionality, it seems these kinds of features are delegated to other more suitable...

...affinity and slower kinetics iGluSnFR3 v857: decreased affinity and faster kinetics which are illustrated... it is selectively activated by glutamate, and tweaking it to improve its responsiveness, they showed ...Improve sensor’s photophysical properties such as kinetics, ∆F/F0, SNR, dynamic range, photostability, quantum...v82) and lime green (v857) variants differing in kinetics, affinity, and ∆F/F0. Introduce the two variants..., GPI, and SGZ, to improve neuronal membrane trafficking and localization. Improved photophysical properties...al…, Supplementary Figure 2.   Improved trafficking and localization through different C-terminal...“Glutamate indicators with improved activation kinetics and localiation for imaging synaptic transmission...

...Scientists are known for asking "What? and "How?" , but they are not always known for asking "Should we?". Scientists...discussed his work with Parkinson's Disease; Hideyuki Okano presented ground-breaking work on spinal cord ...primary Addgene contact for many years, I've been looking forward to seeing how the sharing of his OSKM factors...clinical trials for ALS.   I have an uncle with Parkinson's, a half-brother who is a quadriplegic due to ...techniques are neither fast nor inexpensive.  After talking to numerous poster presenters, it was gratifying... Eric J. Perkins...

...last for several days.” Working with CRISPR Recently Doench has been working on developing CRISPR pooled...not, well, your grade is lower.' ... When you are waking up on a Saturday at like 7 am to go check your ...” Doench has spent most of his research career working on miRNAs and RNAi. He first heard about RNAi at...technology development alongside it.” Doench starting working with David Root in the then RNAi Platform, which...investigate the human genome. Doench says he enjoys working on technology development under the Platform model... but work in true collaboration with scientists asking biological questions. Doench loves that he still... “very much in line with the overall mission of making science easier and more transparent. That’s largely...

...consider when seeking your dream lab. The principal investigator's point of view If you are looking to join...decision. A science trainee will spend 4-7 years working for one person and with a group who all strive ...join a lab, you may be wondering “What is this PI looking for in a new researcher?,” “How do I convince the...typically go through a rotation, which reveals their working habits, personality, and gives them the opportunity...style to spur you to succeed or you might enjoy working closely with others to accomplish your goals. You... You must understand your own style before making this choice.  How big is the lab? While larger labs ... Dr. George Church (Harvard Medical School) for taking the time to speak with us about adding new scientists...

...improve resolution, use a stacking gel on top of the resolving gel. A stacking gel typically has a different...buffer, blocking the membrane  from binding to the antibodies during the stain. The ideal blocking buffer...frequently include suggestions for working concentrations, blocking buffers, and incubation times. If ...interested in making your own knockout line for validation, please view this tutorial from Stuart Orkin's and Daniel...of the proteins in a sample migrate through the stacking gel at the same pace and enter the resolving gel...

...by your library. After blocking for 1 h at room temperature with gentle shaking, gently wick off excess...display little to no homology to anything else, making it difficult to quickly identify possible gene ... overexpression in E. coli, such as pET30a. If working with DNA that is refractory to enzymatic manipulation...the T7 polymerase and a few proteases knocked out making them particularly useful for protein production...harvested dose of cells (preferably in log phase) in blocking solution (around 108 CFU/mL is a good place to...start) and add your membranes to the solution, shaking gently for 1 h at room temperature. This will allow...onto nitrocellulose and carry out the steps from blocking the membrane to probing with cells in order to...

...protein-peptide locking, Mark Howarth’s lab has developed a new tool for peptide-peptide locking. The new technology...light-activated receptor tyrosine kinases Receptor tyrosine kinases (RTKs) are a large class of cell-surface...informational videos, download protocols, and more! Looking for background information on CRISPR technology...Easy-to-use tools to monitor these features were lacking until Roberta Gottlieb and Zhen Yan labs developed...described as “knocksideways” (a British idiom for “taking by surprise”) by Margaret Robinson’s lab, and more...mitochondrial membrane. Sequestration is accomplished by knocking down your endogenous protein of interest and co-expressing... blue and green light-excitable and has faster kinetics than those of other channelrhodopsins. CnChR1,...

...information, making it available to everyone, that creates more breakthrough and innovation. With any kind of open...'s operations. We’re looking for those “what could go wrong scenarios” and making sure we have adequate...than just picking up the phone and having a discussion. By only relying on email and never picking up the... more about Addgene and heard that Addgene was looking for new board members. Paratore: Kristen Darby,...goals were and so I decided to join. I’ve been working with Addgene since 2014 so I’ve been able to watch...

... from Leah Bury of Microscopic Art. I started thinking about this problem when I recently attended the...the transcription of a gene a megabase away, or looking at the histone methylation status at a double-strand...double-strand break that needs repairing. They may be looking at chromatin at the level of nucleosomes, loops... other areas of biological research. If you’re looking for resources that will help you study how chromatin...with Addgene, but you’ll find many great tools by clicking on the links below and visiting their associated...Sharon Dent Archa Fox Elaine Fuchs Kai Ge Robert Kingston Thoru Pederson Phil Sharp Julie Ahringer Alexander... listed here. If you don’t see the gene you’re looking for on this list, please consider searching directly...

...organizations with similar ideas who were also thinking about reproducibility and shared similar desires...Fortunately, the eLife ambassadors group was already thinking about ways to tackle irreproducibility. Benjamin...organizational skills. Meanwhile, Addgene was also thinking about ways to address irreproducibility and thanks... post-workshop surveys, recording results, and tracking material downloads. This was particularly helpful...community. Lessons learned Instead of continuously talking about a problem and potential solutions, take actionable..., but how do you get them to connect? Through networking of course! Thanks to individuals in the group...right time. Be inclusive and seek solutions by working with communities, not for them - Another strength...

...aerosols that are generated (while talking or pipetting) during working in the laminar airflow are also ...providing proper training and guidance to the people working in the lab facility. This can be done by training...talk, laugh, sneeze or cough. Wear gloves while working and discard them after use. Human contact can spread... spread contamination when you are working without gloves or masks. Make sure to change the gloves and...inside the laminar airflow before and while you are working. Perform routine checks for mycoplasma contamination... should be cleaned regularly and dampness of any kind should be avoided. Distilled water provided to maintain...because you are not the first person to face this kind of problem. Educate others when you can and help...

...very small homology regions (5-25 bp) for repair, making it easier to construct targeting vectors. Addgene...copies the sequence from a repair template with flanking sequence homology for error-free DSB repair. NHEJ...MMEJ uses regions with 5-25 bp of microhomology flanking a DSB to repair DNA. The DNA ends are chewed back...figure above, short (5-25 bp) regions of homology flanking a double-stranded break yield precise deletion... is likely to introduce additional nucleotides flanking the insertion. If the DNA ends anneal incorrectly...5’ and 3’ microhomology to the insertion locus flanking the GFP-Puro cassette. Three double stranded breaks...sets of microhomologies (5’ and 3’) to anneal, knocking the GFP-Puro cassette into the locus (see figure...